Dosing regimens for the mobilization of hematopoietic stem and progenitor cells

ABSTRACT

The invention provides compositions and methods useful for mobilizing populations of hematopoietic stem and progenitor cells within a donor, as well as for determining whether samples of mobilized cells are suitable for release for ex vivo expansion and/or therapeutic use. In accordance with the compositions and methods described herein, mobilized hematopoietic stem and progenitor cells can be withdrawn from a donor and administered to a patient for the treatment of various stem cell disorders, including hematopoietic diseases, metabolic disorders, cancers, and autoimmune diseases, among others. In certain embodiments, the compositions and methods described herein lead to the mobilization of a population of CD34dim cells that have immunosuppressive effects and that can reduce the incidence of graft vs. host disease.

CROSS REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of and priority to U.S. applicationSer. No. 15/834,017, filed on Dec. 6, 2017, U.S. Provisional PatentApplication No. 62/596,056, filed on Dec. 7, 2017, U.S. application Ser.No. 16/101,676, filed on Aug. 13, 2018, U.S. Provisional PatentApplication No. 62/753,656, filed on Oct. 31, 2018, and U.S. ProvisionalPatent Application No. 62/773,954, filed on Nov. 30, 2018, thedisclosure of each of which is hereby incorporated by reference in itsentirety.

FIELD OF THE INVENTION

The invention relates to the mobilization of hematopoietic stem andprogenitor cells from a donor, such as a human donor, and to thetreatment of patients suffering from various pathologies, such as blooddiseases, metabolic disorders, cancers, and autoimmune diseases, amongothers.

BACKGROUND OF THE INVENTION

Despite advances in the medicinal arts, there remains a demand fortreating pathologies of the hematopoietic system, such as diseases of aparticular blood cell, metabolic disorders, cancers, and autoimmuneconditions, among others. While hematopoietic stem cells havesignificant therapeutic potential, a limitation that has hindered theiruse in the clinic has been the difficulty associated with releasinghematopoietic stem cells from the bone marrow into the peripheral bloodof a donor, from which the hematopoietic stem cells may be isolated forinfusion into a patient.

A further limitation is that up to 80% of mobilized peripheral blood(mPB) allogeneic recipients will experience graft-versus-host disease(GVHD). Despite the higher levels of CD3+ T cells in mPB grafts comparedto bone marrow transplants, the level of acute GVHD observed followingtransplant of HLA-matched mPB is comparable to HLA-matched bone marrow.One explanation is that G-CSF mobilized grafts contain myeloid-derivedsuppressor cells (MDSCs) possessing potent immunosuppressive propertiescapable of inhibiting T cell proliferation in vitro. The percentage ofMDSCs is variable in grafts mobilized with G-CSF, and clinical datasuggest that patients transplanted with mPB grafts that contain highernumbers of MDSCs may have better outcomes including lower rates of acuteGVHD (Vendramin et al., (2014) BBMT 20(12):2049-2055).

Accordingly, there is currently a need for compositions and methods forpromoting the mobilization of hematopoietic stem and progenitor cells,and particularly for methods of identifying populations of mobilizedcells that are suitable for therapeutic use. There is also a need forcompositions and methods for promoting the mobilization of hematopoieticstem and progenitor cells that consistently produce higher numbers ofMDSCs than do prior art methods.

SUMMARY OF THE INVENTION

The present invention provides compositions and methods for mobilizinghematopoietic stem and progenitor cells in a subject. For example, thesubject may be a hematopoietic stem and progenitor cell donor (i.e., adonor), such as a mammalian donor, and particularly a human donor. Theinvention additionally provides compositions and methods for thetreatment of disorders, such as stem cell disorders, in a patient, suchas a human patient. Using the compositions and methods described herein,a C-X-C chemokine receptor type 2 (CXCR2) agonist, such as Gro-β or avariant thereof, such as a truncated form of Gro-β (e.g., Gro-β T), asdescribed herein, optionally in combination with a C-X-C chemokinereceptor type 4 (CXCR4) antagonist, such as1,1′-[1,4-phenylenebis(methylene)]-bis-1,4,8,11-tetra-azacyclotetradecaneor a variant thereof, may be administered to a subject in amountssufficient to mobilize hematopoietic stem and progenitor cells.Significantly, the compositions and methods described herein may be usedto mobilize hematopoietic stem and progenitor cells from a stem cellniche within a donor, such as a human donor, into the circulatingperipheral blood of the donor while reducing the mobilization of othercells of the hematopoietic lineage, such as white blood cells,neutrophils, lymphocytes, and monocytes. The compositions and methodsdescribed herein thus enable the selective mobilization of hematopoieticstem and progenitor cells in a donor, which may then be isolated from adonor for therapeutic use.

In some embodiments, the hematopoietic stem or progenitor cells may bemobilized from the bone marrow of the donor to the peripheral blood,from which the hematopoietic stem or progenitor cells may be collectedand/or isolated. Upon collection of the mobilized cells, the withdrawnhematopoietic stem or progenitor cells may then be infused into apatient, which may be the donor or another subject, such as a subjectthat is HLA-matched to the donor, for the treatment of one or morepathologies of the hematopoietic system. In some embodiments, thewithdrawn hematopoietic stem or progenitor cells are first expanded exvivo prior to infusion of these cells, and/or progeny thereof, into thepatient. The compositions and methods described herein provide theimportant clinical benefit of enabling the production of populations ofcells that are enriched in hematopoietic stem cells relative to othercell types, such as leukocytes, neutrophils, and monocytes. Thus, thepopulations of mobilized hematopoietic stem and progenitor cellsproduced using the compositions and methods described herein areparticularly suitable for hematopoietic stem cell transplantationtherapy, optionally preceded by ex vivo expansion in order to increasethe quantity of hematopoietic stem and progenitor cells available forinfusion into a patient.

Further, the methods described herein provide the advantage ofinhibiting leukocytosis in the donor. Leukocytosis may lead to adverseeffects such as splenic rupture, renal dysfunction, acute febrilenoninfectious pneumonitis (i.e., pulmonary toxicity), cardiovasculartoxicity (e.g., hypercoagulation, heart attack, dyspnea, angina,arrhythmia, atherosclerotic plaque rupture due to proinflammatoryactions related o high neutrophil counts), neurological disturbances(e.g., blurred vision, headache, and retinal hemorrhage), and sicklecell crisis. See, e.g., D'Souza et al. (2008) Transfusion MedicineReviews 22(4):280-290.

As described herein, hematopoietic stem cells are capable ofdifferentiating into a multitude of cell types in the hematopoieticlineage and can thus be administered to a patient in order to populateor repopulate a cell type that is defective or deficient in the patient.The patient may be one, for example, that is suffering from one or moreblood disorders, such as an autoimmune disease, cancer,hemoglobinopathy, or other hematopoietic pathology, and is therefore inneed of hematopoietic stem cell transplantation. The invention thusprovides methods of treating a variety of hematopoietic conditions, suchas sickle cell anemia, thalassemia, Fanconi anemia, Wiskott-Aldrichsyndrome, adenosine deaminase deficiency-severe combinedimmunodeficiency, metachromatic leukodystrophy, Diamond-Blackfan anemiaand Schwachman-Diamond syndrome, human immunodeficiency virus infection,and acquired immune deficiency syndrome, as well as cancers andautoimmune diseases, among others.

In a first aspect, the invention features a method of mobilizing apopulation of hematopoietic stem cells from the bone marrow of amammalian donor (e.g., a human donor) into peripheral blood, the methodincluding administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to produce a population of cells havinga ratio of CD34⁺ cells to leukocytes of from about 0.0008 to about0.0021 in a sample of peripheral blood of the donor followingadministration of the CXCR2 agonist and CXCR4 antagonist. In someembodiments, the ratio of CD34⁺ cells to leukocytes in the sample may beabout 0.00080, 0.00081, 0.00082, 0.00083, 0.00084, 0.00085, 0.00086,0.00087, 0.00088, 0.00089, 0.00090, 0.00091, 0.00092, 0.00093, 0.00094,0.00095, 0.00096, 0.00097, 0.00098, 0.00099, 0.00100, 0.00101, 0.00102,0.00103, 0.00104, 0.00105, 0.00106, 0.00107, 0.00108, 0.00109, 0.00110,0.00111, 0.00112, 0.00113, 0.00114, 0.00115, 0.00116, 0.00117, 0.00118,0.00119, 0.00120, 0.00121, 0.00122, 0.00123, 0.00124, 0.00125, 0.00126,0.00127, 0.00128, 0.00129, 0.00130, 0.00131, 0.00132, 0.00133, 0.00134,0.00135, 0.00136, 0.00137, 0.00138, 0.00139, 0.00140, 0.00141, 0.00142,0.00143, 0.00144, 0.00145, 0.00146, 0.00147, 0.00148, 0.00149, 0.00150,0.00151, 0.00152, 0.00153, 0.00154, 0.00155, 0.00156, 0.00157, 0.00158,0.00159, 0.00160, 0.00161, 0.00162, 0.00163, 0.00164, 0.00165, 0.00166,0.00167, 0.00168, 0.00169, 0.00170, 0.00171, 0.00172, 0.00173, 0.00174,0.00175, 0.00176, 0.00178, 0.00179, 0.00180, 0.00181, 0.00182, 0.00183,0.00184, 0.00185, 0.00186, 0.00187, 0.00188, 0.00189, 0.00190, 0.00191,0.00192, 0.00193, 0.00194, 0.00195, 0.00196, 0.00197, 0.00198, 0.00199,0.00200, 0.00201, 0.00202, 0.00203, 0.00204, 0.00205, 0.00206, 0.00207,0.00208, 0.00209, 0.00210, 0.00211, 0.00212, 0.00213, 0.00214, 0.00215,0.00216, 0.00217, 0.00218, 0.00219, 0.00220, 0.00221, 0.00222, 0.00223,0.00224, or 0.00225. In some embodiments, the ratio of CD34⁺ cells toleukocytes in the sample is from about 0.0009 to about 0.002, about0.001 to about 0.0019, about 0.0011 to about 0.0018, about 0.0012 toabout 0.0017, about 0.0013 to about 0.0016, or about 0.0014 to about0.0015. In some embodiments, the ratio of CD34⁺ cells to leukocytes inthe sample is from about 0.0010 to about 0.0018, such as a ratio ofhematopoietic stem cells to leukocytes in the sample of about 0.00100,0.00101, 0.00102, 0.00103, 0.00104, 0.00105, 0.00106, 0.00107, 0.00108,0.00109, 0.00110, 0.00111, 0.00112, 0.00113, 0.00114, 0.00115, 0.00116,0.00117, 0.00118, 0.00119, 0.00120, 0.00121, 0.00122, 0.00123, 0.00124,0.00125, 0.00126, 0.00127, 0.00128, 0.00129, 0.00130, 0.00131, 0.00132,0.00133, 0.00134, 0.00135, 0.00136, 0.00137, 0.00138, 0.00139, 0.00140,0.00141, 0.00142, 0.00143, 0.00144, 0.00145, 0.00146, 0.00147, 0.00148,0.00149, 0.00150, 0.00151, 0.00152, 0.00153, 0.00154, 0.00155, 0.00156,0.00157, 0.00158, 0.00159, 0.00160, 0.00161, 0.00162, 0.00163, 0.00164,0.00165, 0.00166, 0.00167, 0.00168, 0.00169, 0.00170, 0.00171, 0.00172,0.00173, 0.00174, 0.00175, 0.00176, 0.00178, 0.00179, or 0.00180. Insome embodiments, the ratio of CD34⁺ cells to leukocytes in the sampleis from about 0.0012 to about 0.0016, such as a ratio of CD34⁺ cells toleukocytes in the sample of about 0.00120, 0.00121, 0.00122, 0.00123,0.00124, 0.00125, 0.00126, 0.00127, 0.00128, 0.00129, 0.00130, 0.00131,0.00132, 0.00133, 0.00134, 0.00135, 0.00136, 0.00137, 0.00138, 0.00139,0.00140, 0.00141, 0.00142, 0.00143, 0.00144, 0.00145, 0.00146, 0.00147,0.00148, 0.00149, 0.00150, 0.00151, 0.00152, 0.00153, 0.00154, 0.00155,0.00156, 0.00157, 0.00158, 0.00159, or 0.00160. In some embodiments, theratio of CD34⁺ cells to leukocytes in the sample is about 0.0014.

In another aspect, the invention features a method of mobilizing apopulation of hematopoietic stem cells from the bone marrow of amammalian donor (e.g., a human donor) into peripheral blood, the methodincluding administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to enrich the peripheral blood of thedonor with CD34⁺ cells relative to leukocytes by a ratio of from about3.40:1 to about 6.90:1 as assessed by comparing a sample of peripheralblood of the donor following administration of the CXCR2 agonist andCXCR4 antagonist to a sample of peripheral blood of the donor prior toadministration of the CXCR2 agonist and CXCR4 antagonist. In someembodiments, the peripheral blood of the donor may be enriched withCD34⁺ cells relative to leukocytes by a ratio of about 3.40:1, 3.45:1,3.50:1, 3.55:1, 3.60:1, 3.65:1, 3.70:1, 3.75:1, 3.80:1, 3.85:1, 3.90:1,3.95:1, 4.00:1, 4.05:1, 4.10:1, 4.15:1, 4.20:1, 4.25:1, 4.30:1, 4.35:1,4.40:1, 4.45:1, 4.50:1, 4.55:1, 4.60:1, 4.65:1, 4.70:1, 4.75:1, 4.80:1,4.85:1, 4.90:1, 4.95:1, 5.00:1, 5.05:1, 5.10:1, 5.15:1, 5.20:1, 5.25:1,5.30:1, 5.35:1, 5.40:1, 5.45:1, 5.50:1, 5.55:1, 5.60:1, 5.65:1, 5.70:1,5.75:1, 5.80:1, 5.85:1, 5.90:1, 5.95:1, 6.00:1, 6.05:1, 6.10:1, 6.15:1,6.20:1, 6.25:1, 6.30:1, 6.35:1, 6.40:1, 6.45:1, 6.50:1, 6.55:1, 6.60:1,6.65:1, 6.70:1, 6.75:1, 6.80:1, 6.85:1, or 6.90:11. In some embodiments,the peripheral blood of the donor is enriched with CD34+ cells relativeto leukocytes by a ratio of from about 3.5:1 to about 6.8:1, about 3.6:1to about 6.7:1, about 3.8:1 to about 6.6:1, about 3.9:1 to about 6.5:1,about 4:1 to about 6.4:1, about 4.1:1 to about 6.3:1, about 4.2:1 toabout 6.2:1, about 4.3:1 to about 6.1:1, about 4.4:1 to about 6:1, about4.5:1 to about 6:1, about 4.6:1 to about 5.9:1, about 4.7:1 to about5.8:1, or about 4.8:1 to about 5.7:1. In some embodiments, theperipheral blood of the donor is enriched with CD34⁺ cells relative toleukocytes by a ratio of about from about 4.0:1 to about 6.0:1, such asa ratio of about 4.00:1, 4.05:1, 4.10:1, 4.15:1, 4.20:1, 4.25:1, 4.30:1,4.35:1, 4.40:1, 4.45:1, 4.50:1, 4.55:1, 4.60:1, 4.65:1, 4.70:1, 4.75:1,4.80:1, 4.85:1, 4.90:1, 4.95:1, 5.00:1, 5.05:1, 5.10:1, 5.15:1, 5.20:1,5.25:1, 5.30:1, 5.35:1, 5.40:1, 5.45:1, 5.50:1, 5.55:1, 5.60:1, 5.65:1,5.70:1, 5.75:1, 5.80:1, 5.85:1, 5.90:1, 5.95:1, or 6.00:1. In someembodiments, the peripheral blood of the donor is enriched with CD34⁺cells relative to leukocytes by a ratio of about from about 4.5:1 toabout 5.5:1, such as a ratio of about 4.50:1, 4.55:1, 4.60:1, 4.65:1,4.70:1, 4.75:1, 4.80:1, 4.85:1, 4.90:1, 4.95:1, 5.00:1, 5.05:1, 5.10:1,5.15:1, 5.20:1, 5.25:1, 5.30:1, 5.35:1, 5.40:1, 5.45:1, or 5.50. In someembodiments, the peripheral blood of the donor is enriched with CD34⁺cells relative to leukocytes by a ratio of about 5.1:1.

In another aspect, the invention features a method of mobilizing apopulation of hematopoietic stem cells from the bone marrow of amammalian donor (e.g., a human donor) into peripheral blood, the methodincluding administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to produce a population of cells havinga density of CD34⁺ cells of at least about 38,000 cells/ml, such as adensity of CD34⁺ cells of from about 38,000 cells/ml to about 100,000cells/ml, about 40,000 cells/ml to about 90,000 cells/nil, about 50,000cells/ml to about 80,000 cells/ml, or about 60,000 cells/ml to about70,000 cells/ml (e.g., about 38,00 cells/ml, 39,000 cells/ml, 40,000cells/ml, 41,000 cells/ml, 42,000 cells/ml, 43,000 cells/ml, 44,000cells/ml, 45,000 cells/nil, 46,000 cells/ml, 47,000 cells/ml, 48,000cells/ml, 49,000 cells/ml, 50,000 cells/ml, 51,000 cells/ml, 52,000cells/ml, 53,000 cells/ml, 54,000 cells/ml, 55,000 cells/ml, 56,000cells/ml, 57,000 cells/ml, 58,000 cells/ml, 59,000 cells/ml, 60,000cells/ml, 61,000 cells/ml, 62,000 cells/ml, 63,000 cells/ml, 64,000cells/ml, 65,000 cells/ml, 66,000 cells/ml, 67,000 cells/ml, 68,000cells/ml, 69,000 cells/ml, 70,000 cells/ml, 71,000 cells/ml, 72,000cells/ml, 73,000 cells/ml, 74,000 cells/ml, 75,000 cells/ml, 76,000cells/ml, 77,000 cells/ml, 78,000 cells/ml, 79,000 cells/ml, 80,000cells/ml, 81,000 cells/ml, 82,000 cells/ml, 83,000 cells/ml, 84,000cells/ml, 85,000 cells/ml, 86,000 cells/ml, 87,000 cells/ml, 88,000cells/ml, 89,000 cells/ml, 90,000 cells/ml, 91,000 cells/ml, 92,000cells/ml, 93,000 cells/ml, 94,000 cells/ml, 95,000 cells/ml, 96,000cells/ml, 97,000 cells/ml, 98,000 cells/ml, 99,000 cells/ml, 100,000cells/ml, or more), and having a density of leukocytes of no more thanabout 5.3×10⁷ cells/ml, such as a density of leukocytes of about 2.3×10⁷cells/ml to about 5.3×10⁷ cells/ml, about 2.5×10⁷ cells/ml to about5.1×10⁷ cells/ml, 2.9×10⁷ cells/ml to about 4.5×10⁷ cells/ml, about3×10⁷ cells/ml to about 4×10⁷ cells/ml (e.g., 5.3×10⁷ cells/ml, 5.2×10⁷cells/ml, 5.1×10⁷ cells/ml, 5×10⁷ cells/ml, 4.9×10⁷ cells/ml, 4.8×10⁷cells/ml, 4.7×10⁷ cells/ml, 4.6×10⁷ cells/ml, 4.5×10⁷ cells/ml, 4.4×10⁷cells/ml, 4.3×10⁷ cells/ml 4.2×10⁷ cells/ml, 4.1×10⁷ cells/ml 4×10⁷cells/ml, 3.9×10⁷ cells/ml, 3.8×10⁷ cells/ml, 3.7×10⁷ cells/ml, 3.6×10⁷cells/ml, 3.5×10⁷ cells/ml, 3.4×10⁷ cells/ml, 3.3×10⁷ cells/ml, 3.2×10⁷cells/ml, 3.1×10⁷ cells/ml, 3×10⁷ cells/ml, 2.9×10⁷ cells/ml, 2.8×10⁷cells/ml, 2.7×10⁷ cells/ml, 2.6×10⁷ cells/ml, 2.5×10⁷ cells/ml, 2.4×10⁷cells/ml, 2.3×10⁷ cells/ml, or less). In some embodiments, the methodincludes administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to produce a population of cells havinga density of CD34⁺ cells of from about 38,000 cells/ml to about 100,000cells/ml, and having a density of leukocytes of from about 2.3×10⁷cells/ml to about 5.3×10⁷ cells/ml. In some embodiments, the methodincludes administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to produce a population of cells havinga density of CD34⁺ cells of from about 40,000 cells/ml to about 80,000cells/ml, and having a density of leukocytes of from about 2.5×10⁷cells/ml to about 5×10⁷ cells/ml. In some embodiments, the methodincludes administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to produce a population of cells havinga density of CD34⁺ cells of from about 50,000 cells/ml to about 90,000cells/ml, and having a density of leukocytes of from about 3×10⁷cells/ml to about 4×10⁷ cells/ml.

In a further aspect, the invention features a method of mobilizing apopulation of hematopoietic stem cells from the bone marrow of amammalian donor (e.g., a human donor) into peripheral blood, the methodincluding administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to produce a population of cells havinga ratio of CD34⁺ cells to neutrophils of from about 0.0018 to about0.0058 in a sample of peripheral blood of the donor followingadministration of the CXCR2 agonist and CXCR4 antagonist. In someembodiments, the ratio of CD34⁺ cells to neutrophils in the sample maybe about 0.00180, 0.00181, 0.00182, 0.00183, 0.00184, 0.00185, 0.00186,0.00187, 0.00188, 0.00189, 0.00190, 0.00191, 0.00192, 0.00193, 0.00194,0.00195, 0.00196, 0.00197, 0.00198, 0.00199, 0.00200, 0.00201, 0.00202,0.00203, 0.00204, 0.00205, 0.00206, 0.00207, 0.00208, 0.00209, 0.00210,0.00211, 0.00212, 0.00213, 0.00214, 0.00215, 0.00216, 0.00217, 0.00218,0.00219, 0.00220, 0.00221, 0.00222, 0.00223, 0.00224, 0.00225, 0.00226,0.00227, 0.00228, 0.00229, 0.00230, 0.00231, 0.00232, 0.00233, 0.00234,0.00235, 0.00236, 0.00237, 0.00238, 0.00239, 0.00240, 0.00241, 0.00242,0.00243, 0.00244, 0.00245, 0.00246, 0.00247, 0.00248, 0.00249, 0.00250,0.00251, 0.00252, 0.00253, 0.00254, 0.00255, 0.00256, 0.00257, 0.00258,0.00259, 0.00260, 0.00261, 0.00262, 0.00263, 0.00264, 0.00265, 0.00266,0.00267, 0.00268, 0.00269, 0.00270, 0.00271, 0.00272, 0.00273, 0.00274,0.00275, 0.00276, 0.00277, 0.00278, 0.00279, 0.00280, 0.00281, 0.00282,0.00283, 0.00284, 0.00285, 0.00286, 0.00287, 0.00288, 0.00289, 0.00290,0.00291, 0.00292, 0.00293, 0.00294, 0.00295, 0.00296, 0.00297, 0.00298,0.00299, 0.00300, 0.00300, 0.00301, 0.00302, 0.00303, 0.00304, 0.00305,0.00306, 0.00307, 0.00308, 0.00309, 0.00310, 0.00311, 0.00312, 0.00313,0.00314, 0.00315, 0.00316, 0.00317, 0.00318, 0.00319, 0.00320, 0.00321,0.00322, 0.00323, 0.00324, 0.00325, 0.00326, 0.00327, 0.00328, 0.00329,0.00330, 0.00331, 0.00332, 0.00333, 0.00334, 0.00335, 0.00336, 0.00337,0.00338, 0.00339, 0.00340, 0.00341, 0.00342, 0.00343, 0.00344, 0.00345,0.00346, 0.00347, 0.00348, 0.00349, 0.00350, 0.00351, 0.00352, 0.00353,0.00354, 0.00355, 0.00356, 0.00357, 0.00358, 0.00359, 0.00360, 0.00361,0.00362, 0.00363, 0.00364, 0.00365, 0.00366, 0.00367, 0.00368, 0.00369,0.00370, 0.00371, 0.00372, 0.00373, 0.00374, 0.00375, 0.00376, 0.00377,0.00378, 0.00379, 0.00380, 0.00381, 0.00382, 0.00383, 0.00384, 0.00385,0.00386, 0.00387, 0.00388, 0.00389, 0.00390, 0.00391, 0.00392, 0.00393,0.00394, 0.00395, 0.00396, 0.00397, 0.00398, 0.00399, 0.00400, 0.00401,0.00402, 0.00403, 0.00404, 0.00405, 0.00406, 0.00407, 0.00408, 0.00409,0.00410, 0.00411, 0.00412, 0.00413, 0.00414, 0.00415, 0.00416, 0.00417,0.00418, 0.00419, 0.00420, 0.00421, 0.00422, 0.00423, 0.00424, 0.00425,0.00426, 0.00427, 0.00428, 0.00429, 0.00430, 0.00431, 0.00432, 0.00433,0.00434, 0.00435, 0.00436, 0.00437, 0.00438, 0.00439, 0.00440, 0.00441,0.00442, 0.00443, 0.00444, 0.00445, 0.00446, 0.00447, 0.00448, 0.00449,0.00450, 0.00451, 0.00452, 0.00453, 0.00454, 0.00455, 0.00456, 0.00457,0.00458, 0.00459, 0.00460, 0.00461, 0.00462, 0.00463, 0.00464, 0.00465,0.00466, 0.00467, 0.00468, 0.00469, 0.00470, 0.00471, 0.00472, 0.00473,0.00474, 0.00475, 0.00476, 0.00477, 0.00478, 0.00479, 0.00480, 0.00481,0.00482, 0.00483, 0.00484, 0.00485, 0.00486, 0.00487, 0.00488, 0.00489,0.00490, 0.00491, 0.00492, 0.00493, 0.00494, 0.00495, 0.00496, 0.00497,0.00498, 0.00499, 0.00500, 0.00501, 0.00502, 0.00503, 0.00504, 0.00505,0.00506, 0.00507, 0.00508, 0.00509, 0.00510, 0.00511, 0.00512, 0.00513,0.00514, 0.00515, 0.00516, 0.00517, 0.00518, 0.00519, 0.00520, 0.00521,0.00522, 0.00523, 0.00524, 0.00525, 0.00526, 0.00527, 0.00528, 0.00529,0.00530, 0.00531, 0.00532, 0.00533, 0.00534, 0.00535, 0.00536, 0.00537,0.00538, 0.00539, 0.00540, 0.00541, 0.00542, 0.00543, 0.00544, 0.00545,0.00546, 0.00547, 0.00548, 0.00549, 0.00550, 0.00551, 0.00552, 0.00553,0.00554, 0.00555, 0.00556, 0.00557, 0.00558, 0.00559, 0.00560, 0.00561,0.00562, 0.00563, 0.00564, 0.00565, 0.00566, 0.00567, 0.00568, 0.00569,0.00570, 0.00571, 0.00572, 0.00573, 0.00574, 0.00575, 0.00576, 0.00577,0.00578, 0.00579, or 0.00580. In some embodiments, the ratio of CD34⁺cells to neutrophils in the sample is from about 0.002 to about 0.0056,about 0.0022 to about 0.0054, about 0.0024 to about 0.0052, about 0.0026to about 0.005, about 0.0028 to about 0.0048, or about 0.003 to about0.0046. In some embodiments, the ratio of CD34⁺ cells to neutrophils inthe sample is from about 0.0026 to about 0.0046, such as a ratio ofCD34⁺ cells to neutrophils in the sample of about 0.00260, 0.00261,0.00262, 0.00263, 0.00264, 0.00265, 0.00266, 0.00267, 0.00268, 0.00269,0.00270, 0.00271, 0.00272, 0.00273, 0.00274, 0.00275, 0.00276, 0.00277,0.00278, 0.00279, 0.00280, 0.00281, 0.00282, 0.00283, 0.00284, 0.00285,0.00286, 0.00287, 0.00288, 0.00289, 0.00290, 0.00291, 0.00292, 0.00293,0.00294, 0.00295, 0.00296, 0.00297, 0.00298, 0.00299, 0.00300, 0.00300,0.00301, 0.00302, 0.00303, 0.00304, 0.00305, 0.00306, 0.00307, 0.00308,0.00309, 0.00310, 0.00311, 0.00312, 0.00313, 0.00314, 0.00315, 0.00316,0.00317, 0.00318, 0.00319, 0.00320, 0.00321, 0.00322, 0.00323, 0.00324,0.00325, 0.00326, 0.00327, 0.00328, 0.00329, 0.00330, 0.00331, 0.00332,0.00333, 0.00334, 0.00335, 0.00336, 0.00337, 0.00338, 0.00339, 0.00340,0.00341, 0.00342, 0.00343, 0.00344, 0.00345, 0.00346, 0.00347, 0.00348,0.00349, 0.00350, 0.00351, 0.00352, 0.00353, 0.00354, 0.00355, 0.00356,0.00357, 0.00358, 0.00359, 0.00360, 0.00361, 0.00362, 0.00363, 0.00364,0.00365, 0.00366, 0.00367, 0.00368, 0.00369, 0.00370, 0.00371, 0.00372,0.00373, 0.00374, 0.00375, 0.00376, 0.00377, 0.00378, 0.00379, 0.00380,0.00381, 0.00382, 0.00383, 0.00384, 0.00385, 0.00386, 0.00387, 0.00388,0.00389, 0.00390, 0.00391, 0.00392, 0.00393, 0.00394, 0.00395, 0.00396,0.00397, 0.00398, 0.00399, 0.00400, 0.00401, 0.00402, 0.00403, 0.00404,0.00405, 0.00406, 0.00407, 0.00408, 0.00409, 0.00410, 0.00411, 0.00412,0.00413, 0.00414, 0.00415, 0.00416, 0.00417, 0.00418, 0.00419, 0.00420,0.00421, 0.00422, 0.00423, 0.00424, 0.00425, 0.00426, 0.00427, 0.00428,0.00429, 0.00430, 0.00431, 0.00432, 0.00433, 0.00434, 0.00435, 0.00436,0.00437, 0.00438, 0.00439, 0.00440, 0.00441, 0.00442, 0.00443, 0.00444,0.00445, 0.00446, 0.00447, 0.00448, 0.00449, 0.00450, 0.00451, 0.00452,0.00453, 0.00454, 0.00455, 0.00456, 0.00457, 0.00458, 0.00459, or0.00460. In some embodiments, the ratio of CD34⁺ cells to neutrophils inthe sample is about 0.0036.

In an additional aspect, the invention features a method of mobilizing apopulation of hematopoietic stem cells from the bone marrow of amammalian donor (e.g., a human donor) into peripheral blood, the methodincluding administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to enrich the peripheral blood of thedonor with CD34⁺ cells relative to neutrophils by a ratio of from about2.1:1 to about 8.1:1 as assessed by comparing a sample of peripheralblood of the donor following administration of the CXCR2 agonist andCXCR4 antagonist to a sample of peripheral blood of the donor prior toadministration of the CXCR2 agonist and CXCR4 antagonist. In someembodiments, the peripheral blood of the donor may be enriched withCD34⁺ cells relative to neutrophils by a ratio of about 2.10:1, 2.15:1,2.20:1, 2.25:1, 2.30:1, 2.35:1, 2.40:1, 2.45:1, 2.50:1, 2.55:1, 2.60:1,2.65:1, 2.70:1, 2.75:1, 2.80:1, 2.85:1, 2.90:1, 2.95:1, 3.00:1, 3.05:1,3.10:1, 3.15:1, 3.20:1, 3.25:1, 3.30:1, 3.35:1, 3.40:1, 3.45:1, 3.50:1,3.55:1, 3.60:1, 3.65:1, 3.70:1, 3.75:1, 3.80:1, 3.85:1, 3.90:1, 3.95:1,4.00:1, 4.05:1, 4.10:1, 4.15:1, 4.20:1, 4.25:1, 4.30:1, 4.35:1, 4.40:1,4.45:1, 4.50:1, 4.55:1, 4.60:1, 4.65:1, 4.70:1, 4.75:1, 4.80:1, 4.85:1,4.90:1, 4.95:1, 5.00:1, 5.05:1, 5.10:1, 5.15:1, 5.20:1, 5.25:1, 5.30:1,5.35:1, 5.40:1, 5.45:1, 5.50:1, 5.55:1, 5.60:1, 5.65:1, 5.70:1, 5.75:1,5.80:1, 5.85:1, 5.90:1, 5.95:1, 6.00:1, 6.05:1, 6.10:1, 6.15:1, 6.20:1,6.25:1, 6.30:1, 6.35:1, 6.40:1, 6.45:1, 6.50:1, 6.55:1, 6.60:1, 6.65:1,6.70:1, 6.75:1, 6.80:1, 6.85:1, 6.90:1, 6.95:1, 7.00:1, 7.05:1, 7.10:1,7.15:1, 7.20:1, 7.25:1, 7.30:1, 7.35:1, 7.40:1, 7.45:1, 7.50:1, 7.55:1,7.60:1, 7.65:1, 7.70:1, 7.75:1, 7.80:1, 7.85:1, 7.90:1, 7.95:1, or8.00:1. In some embodiments, the peripheral blood of the donor isenriched with CD34⁺ cells relative to neutrophils by a ratio of fromabout 2.5:1 to about 7:1, about 2.6:1 to about 6.9:1, about 2.7:1 toabout 6.8:1, about 2.8:1 to about 6.7:1, about 2.9:1 to about 6.6:1,about 3:1 to about 6.5:1, about 3.2:1 to about 6.4:1, about 3.3:1 toabout 6.3:1, about 3.4:1 to about 6.2:1, or about 3.5:1 to about 6.1:1In some embodiments, the peripheral blood of the donor is enriched withCD34⁺ cells relative to neutrophils by a ratio of from about from about5.4:1 to about 7.4:1, such as a ratio of about 5.40:1, 5.45:1, 5.50:1,5.55:1, 5.60:1, 5.65:1, 5.70:1, 5.75:1, 5.80:1, 5.85:1, 5.90:1, 5.95:1,6.00:1, 6.05:1, 6.10:1, 6.15:1, 6.20:1, 6.25:1, 6.30:1, 6.35:1, 6.40:1,6.45:1, 6.50:1, 6.55:1, 6.60:1, 6.65:1, 6.70:1, 6.75:1, 6.80:1, 6.85:1,6.90:1, 6.95:1, 7.00:1, 7.05:1, 7.10:1, 7.15:1, 7.20:1, 7.25:1, 7.30:1,7.35:1, or 7.40:1. In some embodiments, the peripheral blood of thedonor is enriched with CD34⁺ cells relative to neutrophils by a ratio ofabout 6.4:1.

In yet another aspect, the invention features a method of mobilizing apopulation of hematopoietic stem cells from the bone marrow of amammalian donor (e.g., a human donor) into peripheral blood, the methodincluding administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to produce a population of cells havinga ratio of CD34⁺ cells to lymphocytes of from about 0.0021 to about0.0094 in a sample of peripheral blood of the donor followingadministration of the CXCR2 agonist and CXCR4 antagonist. In someembodiments, the ratio of CD34⁺ cells to lymphocytes in the sample maybe about 0.00210, 0.00211, 0.00212, 0.00213, 0.00214, 0.00215, 0.00216,0.00217, 0.00218, 0.00219, 0.00220, 0.00221, 0.00222, 0.00223, 0.00224,0.00225, 0.00226, 0.00227, 0.00228, 0.00229, 0.00230, 0.00231, 0.00232,0.00233, 0.00234, 0.00235, 0.00236, 0.00237, 0.00238, 0.00239, 0.00240,0.00241, 0.00242, 0.00243, 0.00244, 0.00245, 0.00246, 0.00247, 0.00248,0.00249, 0.00250, 0.00251, 0.00252, 0.00253, 0.00254, 0.00255, 0.00256,0.00257, 0.00258, 0.00259, 0.00260, 0.00261, 0.00262, 0.00263, 0.00264,0.00265, 0.00266, 0.00267, 0.00268, 0.00269, 0.00270, 0.00271, 0.00272,0.00273, 0.00274, 0.00275, 0.00276, 0.00277, 0.00278, 0.00279, 0.00280,0.00281, 0.00282, 0.00283, 0.00284, 0.00285, 0.00286, 0.00287, 0.00288,0.00289, 0.00290, 0.00291, 0.00292, 0.00293, 0.00294, 0.00295, 0.00296,0.00297, 0.00298, 0.00299, 0.00300, 0.00300, 0.00301, 0.00302, 0.00303,0.00304, 0.00305, 0.00306, 0.00307, 0.00308, 0.00309, 0.00310, 0.00311,0.00312, 0.00313, 0.00314, 0.00315, 0.00316, 0.00317, 0.00318, 0.00319,0.00320, 0.00321, 0.00322, 0.00323, 0.00324, 0.00325, 0.00326, 0.00327,0.00328, 0.00329, 0.00330, 0.00331, 0.00332, 0.00333, 0.00334, 0.00335,0.00336, 0.00337, 0.00338, 0.00339, 0.00340, 0.00341, 0.00342, 0.00343,0.00344, 0.00345, 0.00346, 0.00347, 0.00348, 0.00349, 0.00350, 0.00351,0.00352, 0.00353, 0.00354, 0.00355, 0.00356, 0.00357, 0.00358, 0.00359,0.00360, 0.00361, 0.00362, 0.00363, 0.00364, 0.00365, 0.00366, 0.00367,0.00368, 0.00369, 0.00370, 0.00371, 0.00372, 0.00373, 0.00374, 0.00375,0.00376, 0.00377, 0.00378, 0.00379, 0.00380, 0.00381, 0.00382, 0.00383,0.00384, 0.00385, 0.00386, 0.00387, 0.00388, 0.00389, 0.00390, 0.00391,0.00392, 0.00393, 0.00394, 0.00395, 0.00396, 0.00397, 0.00398, 0.00399,0.00400, 0.00401, 0.00402, 0.00403, 0.00404, 0.00405, 0.00406, 0.00407,0.00408, 0.00409, 0.00410, 0.00411, 0.00412, 0.00413, 0.00414, 0.00415,0.00416, 0.00417, 0.00418, 0.00419, 0.00420, 0.00421, 0.00422, 0.00423,0.00424, 0.00425, 0.00426, 0.00427, 0.00428, 0.00429, 0.00430, 0.00431,0.00432, 0.00433, 0.00434, 0.00435, 0.00436, 0.00437, 0.00438, 0.00439,0.00440, 0.00441, 0.00442, 0.00443, 0.00444, 0.00445, 0.00446, 0.00447,0.00448, 0.00449, 0.00450, 0.00451, 0.00452, 0.00453, 0.00454, 0.00455,0.00456, 0.00457, 0.00458, 0.00459, 0.00460, 0.00461, 0.00462, 0.00463,0.00464, 0.00465, 0.00466, 0.00467, 0.00468, 0.00469, 0.00470, 0.00471,0.00472, 0.00473, 0.00474, 0.00475, 0.00476, 0.00477, 0.00478, 0.00479,0.00480, 0.00481, 0.00482, 0.00483, 0.00484, 0.00485, 0.00486, 0.00487,0.00488, 0.00489, 0.00490, 0.00491, 0.00492, 0.00493, 0.00494, 0.00495,0.00496, 0.00497, 0.00498, 0.00499, 0.00500, 0.00501, 0.00502, 0.00503,0.00504, 0.00505, 0.00506, 0.00507, 0.00508, 0.00509, 0.00510, 0.00511,0.00512, 0.00513, 0.00514, 0.00515, 0.00516, 0.00517, 0.00518, 0.00519,0.00520, 0.00521, 0.00522, 0.00523, 0.00524, 0.00525, 0.00526, 0.00527,0.00528, 0.00529, 0.00530, 0.00531, 0.00532, 0.00533, 0.00534, 0.00535,0.00536, 0.00537, 0.00538, 0.00539, 0.00540, 0.00541, 0.00542, 0.00543,0.00544, 0.00545, 0.00546, 0.00547, 0.00548, 0.00549, 0.00550, 0.00551,0.00552, 0.00553, 0.00554, 0.00555, 0.00556, 0.00557, 0.00558, 0.00559,0.00560, 0.00561, 0.00562, 0.00563, 0.00564, 0.00565, 0.00566, 0.00567,0.00568, 0.00569, 0.00570, 0.00571, 0.00572, 0.00573, 0.00574, 0.00575,0.00576, 0.00577, 0.00578, 0.00579, 0.00580, 0.00581, 0.00582, 0.00583,0.00584, 0.00585, 0.00586, 0.00587, 0.00588, 0.00589, 0.00590, 0.00591,0.00592, 0.00593, 0.00594, 0.00595, 0.00596, 0.00597, 0.00598, 0.00599,0.00600, 0.00601, 0.00602, 0.00603, 0.00604, 0.00605, 0.00606, 0.00607,0.00608, 0.00609, 0.00610, 0.00611, 0.00612, 0.00613, 0.00614, 0.00615,0.00616, 0.00617, 0.00618, 0.00619, 0.00620, 0.00621, 0.00622, 0.00623,0.00624, 0.00625, 0.00626, 0.00627, 0.00628, 0.00629, 0.00630, 0.00631,0.00632, 0.00633, 0.00634, 0.00635, 0.00636, 0.00637, 0.00638, 0.00639,0.00640, 0.00641, 0.00642, 0.00643, 0.00644, 0.00645, 0.00646, 0.00647,0.00648, 0.00649, 0.00650, 0.00651, 0.00652, 0.00653, 0.00654, 0.00655,0.00656, 0.00657, 0.00658, 0.00659, 0.00660, 0.00661, 0.00662, 0.00663,0.00664, 0.00665, 0.00666, 0.00667, 0.00668, 0.00669, 0.00670, 0.00671,0.00672, 0.00673, 0.00674, 0.00675, 0.00676, 0.00677, 0.00678, 0.00679,0.00680, 0.00681, 0.00682, 0.00683, 0.00684, 0.00685, 0.00686, 0.00687,0.00688, 0.00689, 0.00690, 0.00691, 0.00692, 0.00693, 0.00694, 0.00695,0.00696, 0.00697, 0.00698, 0.00699, 0.00700, 0.00701, 0.00702, 0.00703,0.00704, 0.00705, 0.00706, 0.00707, 0.00708, 0.00709, 0.00710, 0.00711,0.00712, 0.00713, 0.00714, 0.00715, 0.00716, 0.00717, 0.00718, 0.00719,0.00720, 0.00721, 0.00722, 0.00723, 0.00724, 0.00725, 0.00726, 0.00727,0.00728, 0.00729, 0.00730, 0.00731, 0.00732, 0.00733, 0.00734, 0.00735,0.00736, 0.00737, 0.00738, 0.00739, 0.00740, 0.00741, 0.00742, 0.00743,0.00744, 0.00745, 0.00746, 0.00747, 0.00748, 0.00749, 0.00750, 0.00751,0.00752, 0.00753, 0.00754, 0.00755, 0.00756, 0.00757, 0.00758, 0.00759,0.00760, 0.00761, 0.00762, 0.00763, 0.00764, 0.00765, 0.00766, 0.00767,0.00768, 0.00769, 0.00770, 0.00771, 0.00772, 0.00773, 0.00774, 0.00775,0.00776, 0.00777, 0.00778, 0.00779, 0.00780, 0.00781, 0.00782, 0.00783,0.00784, 0.00785, 0.00786, 0.00787, 0.00788, 0.00789, 0.00790, 0.00791,0.00792, 0.00793, 0.00794, 0.00795, 0.00796, 0.00797, 0.00798, 0.00799,0.00800, 0.00801, 0.00802, 0.00803, 0.00804, 0.00805, 0.00806, 0.00807,0.00808, 0.00809, 0.00810, 0.00811, 0.00812, 0.00813, 0.00814, 0.00815,0.00816, 0.00817, 0.00818, 0.00819, 0.00820, 0.00821, 0.00822, 0.00823,0.00824, 0.00825, 0.00826, 0.00827, 0.00828, 0.00829, 0.00830, 0.00831,0.00832, 0.00833, 0.00834, 0.00835, 0.00836, 0.00837, 0.00838, 0.00839,0.00840, 0.00841, 0.00842, 0.00843, 0.00844, 0.00845, 0.00846, 0.00847,0.00848, 0.00849, 0.00850, 0.00851, 0.00852, 0.00853, 0.00854, 0.00855,0.00856, 0.00857, 0.00858, 0.00859, 0.00860, 0.00861, 0.00862, 0.00863,0.00864, 0.00865, 0.00866, 0.00867, 0.00868, 0.00869, 0.00870, 0.00871,0.00872, 0.00873, 0.00874, 0.00875, 0.00876, 0.00877, 0.00878, 0.00879,0.00880, 0.00881, 0.00882, 0.00883, 0.00884, 0.00885, 0.00886, 0.00887,0.00888, 0.00889, 0.00890, 0.00891, 0.00892, 0.00893, 0.00894, 0.00895,0.00896, 0.00897, 0.00898, 0.00899, 0.00900, 0.00901, 0.00902, 0.00903,0.00904, 0.00905, 0.00906, 0.00907, 0.00908, 0.00909, 0.00910, 0.00911,0.00912, 0.00913, 0.00914, 0.00915, 0.00916, 0.00917, 0.00918, 0.00919,0.00920, 0.00921, 0.00922, 0.00923, 0.00924, 0.00925, 0.00926, 0.00927,0.00928, 0.00929, 0.00930, 0.00931, 0.00932, 0.00933, 0.00934, 0.00935,0.00936, 0.00937, 0.00938, 0.00939, or 0.00940. In some embodiments, theratio of CD34⁺ cells to lymphocytes in the sample is from about 0.0022to about 0.0093, about 0.0023 to about 0.0092, about 0.0024 to about0.0091, about 0.003 to about 0.0085, about 0.0035 to about 0.0075, orabout 0.0045 to about 0.0065. In some embodiments, the ratio of CD34⁺cells to lymphocytes in the sample is from about 0.0025 to about 0.0035,such as a ratio of CD34⁺ cells to lymphocytes in the sample of about0.00250, 0.00251, 0.00252, 0.00253, 0.00254, 0.00255, 0.00256, 0.00257,0.00258, 0.00259, 0.00260, 0.00261, 0.00262, 0.00263, 0.00264, 0.00265,0.00266, 0.00267, 0.00268, 0.00269, 0.00270, 0.00271, 0.00272, 0.00273,0.00274, 0.00275, 0.00276, 0.00277, 0.00278, 0.00279, 0.00280, 0.00281,0.00282, 0.00283, 0.00284, 0.00285, 0.00286, 0.00287, 0.00288, 0.00289,0.00290, 0.00291, 0.00292, 0.00293, 0.00294, 0.00295, 0.00296, 0.00297,0.00298, 0.00299, 0.00300, 0.00300, 0.00301, 0.00302, 0.00303, 0.00304,0.00305, 0.00306, 0.00307, 0.00308, 0.00309, 0.00310, 0.00311, 0.00312,0.00313, 0.00314, 0.00315, 0.00316, 0.00317, 0.00318, 0.00319, 0.00320,0.00321, 0.00322, 0.00323, 0.00324, 0.00325, 0.00326, 0.00327, 0.00328,0.00329, 0.00330, 0.00331, 0.00332, 0.00333, 0.00334, 0.00335, 0.00336,0.00337, 0.00338, 0.00339, 0.00340, 0.00341, 0.00342, 0.00343, 0.00344,0.00345, 0.00346, 0.00347, 0.00348, 0.00349, or 0.00350. In someembodiments, the ratio of CD34⁺ cells to lymphocytes in the sample isabout 0.0031.

In an additional aspect, the invention features a method of mobilizing apopulation of hematopoietic stem cells from the bone marrow of amammalian donor (e.g., a human donor) into peripheral blood, the methodincluding administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to enrich the peripheral blood of thedonor with CD34⁺ cells relative to lymphocytes by a ratio of from about4.8:1 to about 8.4:1 as assessed by comparing a sample of peripheralblood of the donor following administration of the CXCR2 agonist andCXCR4 antagonist to a sample of peripheral blood of the donor prior toadministration of the CXCR2 agonist and CXCR4 antagonist. In someembodiments, the peripheral blood of the donor may be enriched withCD34⁺ cells relative to lymphocytes by a ratio of about 4.80:1, 4.85:1,4.90:1, 4.95:1, 5.00:1, 5.05:1, 5.10:1, 5.15:1, 5.20:1, 5.25:1, 5.30:1,5.35:1, 5.40:1, 5.45:1, 5.50:1, 5.55:1, 5.60:1, 5.65:1, 5.70:1, 5.75:1,5.80:1, 5.85:1, 5.90:1, 5.95:1, 6.00:1, 6.05:1, 6.10:1, 6.15:1, 6.20:1,6.25:1, 6.30:1, 6.35:1, 6.40:1, 6.45:1, 6.50:1, 6.55:1, 6.60:1, 6.65:1,6.70:1, 6.75:1, 6.80:1, 6.85:1, 6.90:1, 6.95:1, 7.00:1, 7.05:1, 7.10:1,7.15:1, 7.20:1, 7.25:1, 7.30:1, 7.35:1, 7.40:1, 7.45:1, 7.50:1, 7.55:1,7.60:1, 7.65:1, 7.70:1, 7.75:1, 7.80:1, 7.85:1, 7.90:1, 7.95:1, 8.00:1,8.05:1, 8.10:1, 8.15:1, 8.20:1, 8.25:1, 8.30:1, 8.35:1, or 8.40:1. Insome embodiments, the peripheral blood of the donor is enriched withCD34⁺ cells relative to lymphocytes by a ratio of from about 5:1 toabout 7:1, about 5.5:1 to about 6.5:1, or about 5.2:1 to about 5.7:1. Insome embodiments, the peripheral blood of the donor is enriched withCD34⁺ cells relative to lymphocytes by a ratio of from about 5.0:1 toabout 6.5:1, such as a ratio of about 5.00:1, 5.05:1, 5.10:1, 5.15:1,5.20:1, 5.25:1, 5.30:1, 5.35:1, 5.40:1, 5.45:1, 5.50:1, 5.55:1, 5.60:1,5.65:1, 5.70:1, 5.75:1, 5.80:1, 5.85:1, 5.90:1, 5.95:1, 6.00:1, 6.05:1,6.10:1, 6.15:1, 6.20:1, 6.25:1, 6.30:1, 6.35:1, 6.40:1, 6.45:1, or6.50:1. In some embodiments, the peripheral blood of the donor isenriched with CD34⁺ cells relative to lymphocytes by a ratio of about5.7:1.

In another aspect, the invention features a method of mobilizing apopulation of hematopoietic stem cells from the bone marrow of amammalian donor (e.g., a human donor) into peripheral blood, the methodincluding administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to produce a population of cells havinga density of CD34⁺ cells of at least about 38,000 cells/ml, such as adensity of CD34⁺ cells of from about 38,000 cells/ml to about 100,000cells/ml, about 40,000 cells/ml to about 90,000 cells/ml, about 50,000cells/ml to about 80,000 cells/ml, or about 60,000 cells/ml to about70,000 cells/ml (e.g., about 38,00 cells/ml, 39,000 cells/ml, 40,000cells/ml, 41,000 cells/ml, 42,000 cells/ml, 43,000 cells/ml, 44,000cells/ml, 45,000 cells/ml, 46,000 cells/ml, 47,000 cells/ml, 48,000cells/ml, 49,000 cells/ml, 50,000 cells/ml, 51,000 cells/ml, 52,000cells/ml, 53,000 cells/ml, 54,000 cells/ml, 55,000 cells/ml, 56,000cells/ml, 57,000 cells/ml, 58,000 cells/ml, 59,000 cells/ml, 60,000cells/ml, 61,000 cells/ml, 62,000 cells/ml, 63,000 cells/ml, 64,000cells/ml, 65,000 cells/ml, 66,000 cells/ml, 67,000 cells/ml, 68,000cells/ml, 69,000 cells/ml, 70,000 cells/ml, 71,000 cells/ml, 72,000cells/ml, 73,000 cells/ml, 74,000 cells/ml, 75,000 cells/ml, 76,000cells/ml, 77,000 cells/ml, 78,000 cells/ml, 79,000 cells/ml, 80,000cells/ml, 81,000 cells/ml, 82,000 cells/ml, 83,000 cells/ml, 84,000cells/ml, 85,000 cells/ml, 86,000 cells/ml, 87,000 cells/ml, 88,000cells/ml, 89,000 cells/ml, 90,000 cells/ml, 91,000 cells/ml, 92,000cells/ml, 93,000 cells/ml, 94,000 cells/ml, 95,000 cells/ml, 96,000cells/ml, 97,000 cells/ml, 98,000 cells/ml, 99,000 cells/ml, 100,000cells/ml, or more), and having a density of lymphocytes of no more thanabout 2.4×10⁷ cells/ml, such as a density of lymphocytes of about 1×10⁷cells/ml to about 2.3×10⁷ cells/ml, about 1.3×10⁷ cells/ml to about2.1×10⁷ cells/ml, or about 1.5×10⁷ cells/ml to about 1.9×10⁷ cells/ml(e.g., about 2.4×10⁷ cells/ml, 2.3×10⁷ cells/ml, 2.2×10⁷ cells/ml,2.1×10⁷ cells/ml, 2×10⁷ cells/ml, 1.9×10⁷ cells/ml, 1.8×10⁷ cells/ml,1.7×10⁷ cells/ml, 1.6×10⁷ cells/ml, 1.5×10⁷ cells/ml 1.4×10⁷ cells/ml,1.3×10⁷ cells/ml, 1.2×10⁷ cells/ml, 1.1×10⁷ cells/ml, 1×10⁷ cells/ml, orless, 0.9×10⁷ cells/ml, 0.8×10⁷ cells/ml, or less). In some embodiments,the method includes administering to the donor a CXCR2 agonist and aCXCR4 antagonist in amounts sufficient to produce a population of cellshaving a density of CD34⁺ cells of from about 38,000 cells/ml to about100,000 cells/ml, and having a density of lymphocytes of from about1×10⁷ cells/ml to about 2.3×10⁷ cells/ml. In some embodiments, themethod includes administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to produce a population of cells havinga density of CD34⁺ cells of from about 40,000 cells/ml to about 80,000cells/ml, and having a density of lymphocytes of from about 1.3×10⁷cells/ml to about 2.3×10⁷ cells/ml. In some embodiments, the methodincludes administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to produce a population of cells havinga density of CD34⁺ cells of from about 50,000 cells/ml to about 90,000cells/ml, and having a density of lymphocytes of from about 1.5×10⁷cells/ml to about 2×10⁷ cells/ml.

In another aspect, the invention features a method of mobilizing apopulation of hematopoietic stem cells from the bone marrow of amammalian donor (e.g., a human donor) into peripheral blood, the methodincluding administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to produce a population of cells havinga ratio of CD34⁺ cells to monocytes of from about 0.0071 to about 0.0174in a sample of peripheral blood of the donor following administration ofthe CXCR2 agonist. In some embodiments, the ratio of CD34⁺ cells tomonocytes in the sample may be about 0.00710, 0.00711, 0.00712, 0.00713,0.00714, 0.00715, 0.00716, 0.00717, 0.00718, 0.00719, 0.00720, 0.00721,0.00722, 0.00723, 0.00724, 0.00725, 0.00726, 0.00727, 0.00728, 0.00729,0.00730, 0.00731, 0.00732, 0.00733, 0.00734, 0.00735, 0.00736, 0.00737,0.00738, 0.00739, 0.00740, 0.00741, 0.00742, 0.00743, 0.00744, 0.00745,0.00746, 0.00747, 0.00748, 0.00749, 0.00750, 0.00751, 0.00752, 0.00753,0.00754, 0.00755, 0.00756, 0.00757, 0.00758, 0.00759, 0.00760, 0.00761,0.00762, 0.00763, 0.00764, 0.00765, 0.00766, 0.00767, 0.00768, 0.00769,0.00770, 0.00771, 0.00772, 0.00773, 0.00774, 0.00775, 0.00776, 0.00777,0.00778, 0.00779, 0.00780, 0.00781, 0.00782, 0.00783, 0.00784, 0.00785,0.00786, 0.00787, 0.00788, 0.00789, 0.00790, 0.00791, 0.00792, 0.00793,0.00794, 0.00795, 0.00796, 0.00797, 0.00798, 0.00799, 0.00800, 0.00801,0.00802, 0.00803, 0.00804, 0.00805, 0.00806, 0.00807, 0.00808, 0.00809,0.00810, 0.00811, 0.00812, 0.00813, 0.00814, 0.00815, 0.00816, 0.00817,0.00818, 0.00819, 0.00820, 0.00821, 0.00822, 0.00823, 0.00824, 0.00825,0.00826, 0.00827, 0.00828, 0.00829, 0.00830, 0.00831, 0.00832, 0.00833,0.00834, 0.00835, 0.00836, 0.00837, 0.00838, 0.00839, 0.00840, 0.00841,0.00842, 0.00843, 0.00844, 0.00845, 0.00846, 0.00847, 0.00848, 0.00849,0.00850, 0.00851, 0.00852, 0.00853, 0.00854, 0.00855, 0.00856, 0.00857,0.00858, 0.00859, 0.00860, 0.00861, 0.00862, 0.00863, 0.00864, 0.00865,0.00866, 0.00867, 0.00868, 0.00869, 0.00870, 0.00871, 0.00872, 0.00873,0.00874, 0.00875, 0.00876, 0.00877, 0.00878, 0.00879, 0.00880, 0.00881,0.00882, 0.00883, 0.00884, 0.00885, 0.00886, 0.00887, 0.00888, 0.00889,0.00890, 0.00891, 0.00892, 0.00893, 0.00894, 0.00895, 0.00896, 0.00897,0.00898, 0.00899, 0.00900, 0.00901, 0.00902, 0.00903, 0.00904, 0.00905,0.00906, 0.00907, 0.00908, 0.00909, 0.00910, 0.00911, 0.00912, 0.00913,0.00914, 0.00915, 0.00916, 0.00917, 0.00918, 0.00919, 0.00920, 0.00921,0.00922, 0.00923, 0.00924, 0.00925, 0.00926, 0.00927, 0.00928, 0.00929,0.00930, 0.00931, 0.00932, 0.00933, 0.00934, 0.00935, 0.00936, 0.00937,0.00938, 0.00939, 0.00940, 0.00941, 0.00942, 0.00943, 0.00944, 0.00945,0.00946, 0.00947, 0.00948, 0.00949, 0.00950, 0.00951, 0.00952, 0.00953,0.00954, 0.00955, 0.00956, 0.00957, 0.00958, 0.00959, 0.00960, 0.00961,0.00962, 0.00963, 0.00964, 0.00965, 0.00966, 0.00967, 0.00968, 0.00969,0.00970, 0.00971, 0.00972, 0.00973, 0.00974, 0.00975, 0.00976, 0.00977,0.00978, 0.00979, 0.00980, 0.00981, 0.00982, 0.00983, 0.00984, 0.00985,0.00986, 0.00987, 0.00988, 0.00989, 0.00990, 0.00991, 0.00992, 0.00993,0.00994, 0.00995, 0.00996, 0.00997, 0.00998, 0.00999, 0.0100, 0.0101,0.0103, 0.0104, 0.0105, 0.0106, 0.0107, 0.0108, 0.0109, 0.0110, 0.0111,0.0112, 0.0113, 0.0114, 0.0115, 0.0116, 0.0117, 0.0118, 0.0119, 0.0120,0.0121, 0.0122, 0.0123, 0.0124, 0.0125, 0.0126, 0.0127, 0.0128, 0.0129,0.0130, 0.0131, 0.0132, 0.0133, 0.0134, 0.0135, 0.0136, 0.0137, 0.0138,0.0139, 0.0140, 0.0141, 0.0142, 0.0143, 0.0144, 0.0145, 0.0146, 0.0147,0.0148, 0.0149, 0.0150, 0.0151, 0.0152, 0.0153, 0.0154, 0.0155, 0.0156,0.0157, 0.0158, 0.0159, 0.0160, 0.0161, 0.0162, 0.0163, 0.0164, 0.0165,0.0166, 0.0167, 0.0168, 0.0169, 0.0170, 0.0171, 0.0172, 0.0173, or0.0174. In some embodiments, the ratio of CD34⁺ cells to monocytes inthe sample is from 0.008 to about 0.016, about 0.009 to about 0.015,about 0.01 to about 0.014, or about 0.011 to about 0.013. In someembodiments, the ratio of CD34⁺ cells to monocytes in the sample is fromabout 0.0100 to about 0.0140, such as a ratio of CD34⁺ cells tomonocytes in the sample of about 0.0100, 0.0101, 0.0103, 0.0104, 0.0105,0.0106, 0.0107, 0.0108, 0.0109, 0.0110, 0.0111, 0.0112, 0.0113, 0.0114,0.0115, 0.0116, 0.0117, 0.0118, 0.0119, 0.0120, 0.0121, 0.0122, 0.0123,0.0124, 0.0125, 0.0126, 0.0127, 0.0128, 0.0129, 0.0130, 0.0131, 0.0132,0.0133, 0.0134, 0.0135, 0.0136, 0.0137, 0.0138, 0.0139, or 0.0140. Insome embodiments, the ratio of CD34⁺ cells to monocytes in the sample isabout 0.0118.

In an additional aspect, the invention features a method of mobilizing apopulation of hematopoietic stem cells from the bone marrow of amammalian donor (e.g., a human donor) into peripheral blood, the methodincluding administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to produce a population of cells havinga density of CD34⁺ cells of at least about 38,000 cells/ml, such as adensity of CD34⁺ cells of from about 38,000 cells/ml to about 100,000cells/ml, about 40,000 cells/ml to about 90,000 cells/ml, about 50,000cells/ml to about 80,000 cells/ml, or about 60,000 cells/ml to about70,000 cells/ml (e.g., about 38,00 cells/ml, 39,000 cells/ml, 40,000cells/ml, 41,000 cells/ml, 42,000 cells/ml, 43,000 cells/ml, 44,000cells/ml, 45,000 cells/ml, 46,000 cells/ml, 47,000 cells/ml, 48,000cells/ml, 49,000 cells/ml, 50,000 cells/ml, 51,000 cells/ml, 52,000cells/ml, 53,000 cells/ml, 54,000 cells/ml, 55,000 cells/ml, 56,000cells/ml, 57,000 cells/ml, 58,000 cells/ml, 59,000 cells/ml, 60,000cells/ml, 61,000 cells/ml, 62,000 cells/ml, 63,000 cells/ml, 64,000cells/ml, 65,000 cells/ml, 66,000 cells/ml, 67,000 cells/ml, 68,000cells/ml, 69,000 cells/ml, 70,000 cells/ml, 71,000 cells/ml, 72,000cells/ml, 73,000 cells/ml, 74,000 cells/ml, 75,000 cells/ml, 76,000cells/ml, 77,000 cells/ml, 78,000 cells/ml, 79,000 cells/ml, 80,000cells/ml, 81,000 cells/ml, 82,000 cells/ml, 83,000 cells/ml, 84,000cells/ml, 85,000 cells/ml, 86,000 cells/ml, 87,000 cells/ml, 88,000cells/ml, 89,000 cells/ml, 90,000 cells/ml, 91,000 cells/ml, 92,000cells/ml, 93,000 cells/ml, 94,000 cells/ml, 95,000 cells/ml, 96,000cells/ml, 97,000 cells/ml, 98,000 cells/ml, 99,000 cells/ml, 100,000cells/ml, or more), and having a density of monocytes of no more thanabout 6×10⁶ cells/ml, such as a density of monocytes of from 3.4×10⁶cells/ml to about 5.9×10⁶ cells/ml, about 3.5×10⁶ cells/ml to about5.7×10⁶ cells/ml, or about 4×10⁶ cells/ml to about 5×10⁶ cells/ml (e.g.,5.9×10⁶ cells/ml, 5.8×10⁶ cells/ml, 5.7×10⁶ cells/ml, 5.6×10⁶ cells/ml,5.5×10⁶ cells/ml, 5.4×10⁶ cells/ml, 5.3×10⁶ cells/ml, 5.2×10⁶ cells/ml,5.1×10⁶ cells/ml, 5×10⁶ cells/ml, 4.9×10⁶ cells/ml, 4.8×10⁶ cells/ml,4.7×10⁶ cells/ml, 4.6×10⁶ cells/ml, 4.5×10⁶ cells/ml, 4.4×10⁶ cells/ml,4.3×10⁶ cells/ml, 4.2×10⁶ cells/ml, 4.1×10⁶ cells/ml, 4×10⁶ cells/ml,3.9×10⁶ cells/ml, 3.8×10⁶ cells/ml, 3.7×10⁶ cells/ml, 3.6×10⁶ cells/ml,3.5×10⁶ cells/ml, 3.4×10⁶ cells/ml, or less). In some embodiments, themethod includes administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to produce a population of cells havinga density of CD34⁺ cells of from about 38,000 cells/ml to about 100,000cells/ml, and having a density of monocytes of from about 3.4×10⁶cells/ml to about 6×10⁶ cells/ml. In some embodiments, the methodincludes administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to produce a population of cells havinga density of CD34⁺ cells of from about 40,000 cells/ml to about 80,000cells/ml, and having a density of monocytes of from about 4×10⁶ cells/mlto about 5.5×10⁶ cells/ml. In some embodiments, the method includesadministering to the donor a CXCR2 agonist and a CXCR4 antagonist inamounts sufficient to produce a population of cells having a density ofCD34⁺ cells of from about 50,000 cells/ml to about 90,000 cells/ml, andhaving a density of monocytes of from about 4×10⁶ cells/ml to about5×10⁶ cells/ml.

In another aspect, the invention features a method of mobilizing apopulation of hematopoietic stem cells from the bone marrow of amammalian donor (e.g., a human donor) into peripheral blood, the methodincluding administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to enrich the peripheral blood of thedonor with CD34⁺ cells relative to monocytes by a ratio of from about1.1:1 to about 2.3:1 as assessed by comparing a sample of peripheralblood of the donor following administration of the CXCR2 agonist andCXCR4 antagonist to a sample of peripheral blood of the donor prior toadministration of the CXCR2 agonist and CXCR4 antagonist. In someembodiments, the peripheral blood of the donor may be enriched withCD34⁺ cells relative to monocytes by a ratio of about 1.10:1, 1.15:1,1.20:1, 1.25:1, 1.30:1, 1.35:1, 1.40:1, 1.45:1, 1.50:1, 1.55:1, 1.60:1,1.65:1, 1.70:1, 1.75:1, 1.80:1, 1.85:1, 1.90:1, 1.95:1, 2.00:1, 2.05:1,2.10:1, 2.15:1, 2.20:1, 2.25:1, or 2.30:1. In some embodiments, theperipheral blood of the donor is enriched with CD34⁺ cells relative tomonocytes by a ratio of about from about 1.3:1 to about 1.9:1, such as aratio of about 1.30:1, 1.35:1, 1.40:1, 1.45:1, 1.50:1, 1.55:1, 1.60:1,1.65:1, 1.70:1, 1.75:1, 1.80:1, 1.85:1, or 1.90:1.

In another aspect, the invention features a method of mobilizing apopulation of hematopoietic stem cells from the bone marrow of amammalian donor (e.g., a human donor) into peripheral blood, the methodincluding administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to produce a population of cells havinga frequency of CD34⁺ cells of from about 0.051% to about 0.140% in asample of peripheral blood of the donor following administration of theCXCR2 agonist and CXCR4 antagonist. In some embodiments, the populationof cells may have a frequency of CD34⁺ cells of about 0.051%, 0.052%,0.053%, 0.054%, 0.055%, 0.056%, 0.057%, 0.058%, 0.059%, 0.060%, 0.061%,0.062%, 0.063%, 0.064%, 0.065%, 0.066%, 0.067%, 0.068%, 0.069%, 0.070%,0.071%, 0.072%, 0.073%, 0.074%, 0.075%, 0.076%, 0.077%, 0.078%, 0.079%,0.080%, 0.081%, 0.082%, 0.083%, 0.084%, 0.085%, 0.086%, 0.087%, 0.088%,0.089%, 0.090%, 0.091%, 0.092%, 0.093%, 0.094%, 0.095%, 0.096%, 0.097%,0.098%, 0.099%, 0.100%, 0.101%, 0.102%, 0.103%, 0.104%, 0.105%, 0.106%,0.107%, 0.108%, 0.109%, 0.110%, 0.111%, 0.112%, 0.113%, 0.114%, 0.115%,0.116%, 0.117%, 0.118%, 0.119%, 0.120%, 0.121%, 0.122%, 0.123%, 0.124%,0.125%, 0.126%, 0.127%, 0.128%, 0.129%, 0.130%, 0.131%, 0.132%, 0.133%,0.134%, 0.135%, 0.136%, 0.137%, 0.138%, 0.139%, or 0.140%. In someembodiments, the population of cells has a frequency of CD34⁺ cells offrom about 0.050% to about 0.120%, about 0.060% to about 0.110%, orabout 0.080% to about 0.100%. In some embodiments, the population ofcells has a frequency of CD34⁺ cells of from about 0.080% to about0.120%, such as a frequency of hematopoietic stem cells of about 0.080%,0.081%, 0.082%, 0.083%, 0.084%, 0.085%, 0.086%, 0.087%, 0.088%, 0.089%,0.090%, 0.091%, 0.092%, 0.093%, 0.094%, 0.095%, 0.096%, 0.097%, 0.098%,0.099%, 0.100%, 0.101%, 0.102%, 0.103%, 0.104%, 0.105%, 0.106%, 0.107%,0.108%, 0.109%, 0.110%, 0.111%, 0.112%, 0.113%, 0.114%, 0.115%, 0.116%,0.117%, 0.118%, 0.119%, or 0.120%. In some embodiments, the populationof cells has a frequency of CD34⁺ cells of about 0.097%.

In an additional aspect, the invention features a method of mobilizing apopulation of hematopoietic stem cells from the bone marrow of amammalian donor (e.g., a human donor) into peripheral blood, the methodincluding administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to induce an increase in the frequencyof CD34⁺ cells in the peripheral blood of the donor by at least 3-foldas assessed by comparing a sample of peripheral blood of the donorfollowing administration of the CXCR2 agonist and CXCR4 antagonist to asample of peripheral blood of the donor prior to administration of theCXCR2 agonist and CXCR4 antagonist (e.g., by from about 3.4-fold toabout 7.1-fold, such as by about 3.4-fold, 3.5-fold, 3.6-fold, 3.7-fold,3.8-fold, 3.9-fold, 4.0-fold, 4.1-fold, 4.2-fold, 4.3-fold, 4.4-fold,4.5-fold, 4.6-fold, 4.7-fold, 4.8-fold, 4.9-fold, 5.0-fold, 5.1-fold,5.2-fold, 5.3-fold, 5.4-fold, 5.5-fold, 5.6-fold, 5.7-fold, 5.8-fold,5.9-fold, 6.0-fold, 6.1-fold, 6.2-fold, 6.3-fold, 6.4-fold, 6.5-fold,6.6-fold, 6.7-fold, 6.8-fold, 6.9-fold, 7.0-fold, or 7.1-fold. In someembodiments, the frequency of CD34⁺ cells in the peripheral blood of thedonor is increased by from about 4-fold to about 7-fold, about 4.5-foldto about 6.5-fold, or about 5-fold to about 6-fold followingadministration of the CXCR2 agonist and CXCR4 antagonist. In someembodiments, the frequency of CD34⁺ cells in the peripheral blood of thedonor is increased by from about 4.0-fold to about 6.0-fold followingadministration of the CXCR2 agonist and CXCR4 antagonist, such as byabout 4.0-fold, 4.1-fold, 4.2-fold, 4.3-fold, 4.4-fold, 4.5-fold,4.6-fold, 4.7-fold, 4.8-fold, 4.9-fold, 5.0-fold, 5.1-fold, 5.2-fold,5.3-fold, 5.4-fold, 5.5-fold, 5.6-fold, 5.7-fold, 5.8-fold, 5.9-fold, or6.0-fold. In some embodiments, the frequency of CD34⁺ cells in theperipheral blood of the donor is increased by about 4.8-fold.

In a further aspect, the invention features a method of mobilizing apopulation of hematopoietic stem cells from the bone marrow of amammalian donor (e.g., a human donor) into peripheral blood, the methodincluding administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to produce a population of cells havinga ratio of CD34⁺ CD90⁺ CD45RA⁻ cells to leukocytes of from about 0.0003to about 0.0016 in a sample of peripheral blood of the donor followingadministration of the CXCR2 agonist and CXCR4 antagonist. In someembodiments, the ratio of CD34⁺ CD90⁺ CD45RA⁻ cells to leukocytes in thesample may be about 0.00030, 0.00031, 0.00032, 0.00033, 0.00034,0.00035, 0.00036, 0.00037, 0.00038, 0.00039, 0.00040, 0.00041, 0.00042,0.00043, 0.00044, 0.00045, 0.00046, 0.00047, 0.00048, 0.00049, 0.00050,0.00051, 0.00052, 0.00053, 0.00054, 0.00055, 0.00056, 0.00057, 0.00058,0.00059, 0.00060, 0.00061, 0.00062, 0.00063, 0.00064, 0.00065, 0.00066,0.00067, 0.00068, 0.00069, 0.00070, 0.00071, 0.00072, 0.00073, 0.00074,0.00075, 0.00076, 0.00077, 0.00078, 0.00079, 0.00080, 0.00081, 0.00082,0.00083, 0.00084, 0.00085, 0.00086, 0.00087, 0.00088, 0.00089, 0.00090,0.00091, 0.00092, 0.00093, 0.00094, 0.00095, 0.00096, 0.00097, 0.00098,0.00099, 0.00100, 0.00101, 0.00102, 0.00103, 0.00104, 0.00105, 0.00106,0.00107, 0.00108, 0.00109, 0.00110, 0.00111, 0.00112, 0.00113, 0.00114,0.00115, 0.00116, 0.00117, 0.00118, 0.00119, 0.00120, 0.00121, 0.00122,0.00123, 0.00124, 0.00125, 0.00126, 0.00127, 0.00128, 0.00129, 0.00130,0.00131, 0.00132, 0.00133, 0.00134, 0.00135, 0.00136, 0.00137, 0.00138,0.00139, 0.00140, 0.00141, 0.00142, 0.00143, 0.00144, 0.00145, 0.00146,0.00147, 0.00148, 0.00149, 0.00150, 0.00151, 0.00152, 0.00153, 0.00154,0.00155, 0.00156, 0.00157, 0.00158, 0.00159, or 0.00160. In someembodiments, the ratio of CD34⁺ CD90⁺ CD45RA⁻ cells to leukocytes in thesample is from about 0.0008 to about 0.0010. In some embodiments, theratio of CD34⁺ CD90⁺ CD45RA⁻ cells to leukocytes in the sample is fromabout 0.0006 to about 0.0012, such as a ratio of hematopoietic stemcells to leukocytes in the sample of about 0.00060, 0.00061, 0.00062,0.00063, 0.00064, 0.00065, 0.00066, 0.00067, 0.00068, 0.00069, 0.00070,0.00071, 0.00072, 0.00073, 0.00074, 0.00075, 0.00076, 0.00077, 0.00078,0.00079, 0.00080, 0.00081, 0.00082, 0.00083, 0.00084, 0.00085, 0.00086,0.00087, 0.00088, 0.00089, 0.00090, 0.00091, 0.00092, 0.00093, 0.00094,0.00095, 0.00096, 0.00097, 0.00098, 0.00099, 0.00100, 0.00101, 0.00102,0.00103, 0.00104, 0.00105, 0.00106, 0.00107, 0.00108, 0.00109, 0.00110,0.00111, 0.00112, 0.00113, 0.00114, 0.00115, 0.00116, 0.00117, 0.00118,0.00119, or 0.00120. In some embodiments, the ratio of CD34⁺ CD90⁺CD45RA⁻ cells to leukocytes in the sample is about 0.0009.

In another aspect, the invention features a method of mobilizing apopulation of hematopoietic stem cells from the bone marrow of amammalian donor (e.g., a human donor) into peripheral blood, the methodincluding administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to enrich the peripheral blood of thedonor with CD34⁺ CD90⁺ CD45RA⁻ cells relative to leukocytes by a ratioof from about 5.5:1 to about 26.9:1 as assessed by comparing a sample ofperipheral blood of the donor following administration of the CXCR2agonist and CXCR4 antagonist to a sample of peripheral blood of thedonor prior to administration of the CXCR2 agonist and CXCR4 antagonist.In some embodiments, the peripheral blood of the donor may be enrichedwith CD34⁺ CD90⁺ CD45RA⁻ cells relative to leukocytes by a ratio ofabout 5.50:1, 5.55:1, 5.60:1, 5.65:1, 5.70:1, 5.75:1, 5.80:1, 5.85:1,5.90:1, 5.95:1, 6.00:1, 6.05:1, 6.10:1, 6.15:1, 6.20:1, 6.25:1, 6.30:1,6.35:1, 6.40:1, 6.45:1, 6.50:1, 6.55:1, 6.60:1, 6.65:1, 6.70:1, 6.75:1,6.80:1, 6.85:1, 6.90:1, 6.95:1, 7.00:1, 7.05:1, 7.10:1, 7.15:1, 7.20:1,7.25:1, 7.30:1, 7.35:1, 7.40:1, 7.45:1, 7.50:1, 7.55:1, 7.60:1, 7.65:1,7.70:1, 7.75:1, 7.80:1, 7.85:1, 7.90:1, 7.95:1, 8.00:1, 8.05:1, 8.10:1,8.15:1, 8.20:1, 8.25:1, 8.30:1, 8.35:1, 8.40:1, 8.45:1, 8.50:1, 8.55:1,8.60:1, 8.65:1, 8.70:1, 8.75:1, 8.80:1, 8.85:1, 8.90:1, 8.95:1, 9.00:1,9.05:1, 9.10:1, 9.15:1, 9.20:1, 9.25:1, 9.30:1, 9.35:1, 9.40:1, 9.45:1,9.50:1, 9.55:1, 9.60:1, 9.65:1, 9.70:1, 9.75:1, 9.80:1, 9.85:1, 9.90:1,9.95:1, 10.00:1, 10.05:1, 10.10:1, 10.15:1, 10.20:1, 10.25:1, 10.30:1,10.35:1, 10.40:1, 10.45:1, 10.50:1, 10.55:1, 10.60:1, 10.65:1, 10.70:1,10.75:1, 10.80:1, 10.85:1, 10.90:1, 10.95:1, 11.00:1, 11.05:1, 11.10:1,11.15:1, 11.20:1, 11.25:1, 11.30:1, 11.35:1, 11.40:1, 11.45:1, 11.50:1,11.55:1, 11.60:1, 11.65:1, 11.70:1, 11.75:1, 11.80:1, 11.85:1, 11.90:1,11.95:1, 12.00:1, 12.05:1, 12.10:1, 12.15:1, 12.20:1, 12.25:1, 12.30:1,12.35:1, 12.40:1, 12.45:1, 12.50:1, 12.55:1, 12.60:1, 12.65:1, 12.70:1,12.75:1, 12.80:1, 12.85:1, 12.90:1, 12.95:1, 13.00:1, 13.05:1, 13.10:1,13.15:1, 13.20:1, 13.25:1, 13.30:1, 13.35:1, 13.40:1, 13.45:1, 13.50:1,13.55:1, 13.60:1, 13.65:1, 13.70:1, 13.75:1, 13.80:1, 13.85:1, 13.90:1,13.95:1, 14.00:1, 14.05:1, 14.10:1, 14.15:1, 14.20:1, 14.25:1, 14.30:1,14.35:1, 14.40:1, 14.45:1, 14.50:1, 14.55:1, 14.60:1, 14.65:1, 14.70:1,14.75:1, 14.80:1, 14.85:1, 14.90:1, 14.95:1, 15.00:1, 15.05:1, 15.10:1,15.15:1, 15.20:1, 15.25:1, 15.30:1, 15.35:1, 15.40:1, 15.45:1, 15.50:1,15.55:1, 15.60:1, 15.65:1, 15.70:1, 15.75:1, 15.80:1, 15.85:1, 15.90:1,15.95:1, 16.00:1, 16.05:1, 16.10:1, 16.15:1, 16.20:1, 16.25:1, 16.30:1,16.35:1, 16.40:1, 16.45:1, 16.50:1, 16.55:1, 16.60:1, 16.65:1, 16.70:1,16.75:1, 16.80:1, 16.85:1, 16.90:1, 16.95:1, 17.00:1, 17.05:1, 17.10:1,17.15:1, 17.20:1, 17.25:1, 17.30:1, 17.35:1, 17.40:1, 17.45:1, 17.50:1,17.55:1, 17.60:1, 17.65:1, 17.70:1, 17.75:1, 17.80:1, 17.85:1, 17.90:1,17.95:1, 18.00:1, 18.05:1, 18.10:1, 18.15:1, 18.20:1, 18.25:1, 18.30:1,18.35:1, 18.40:1, 18.45:1, 18.50:1, 18.55:1, 18.60:1, 18.65:1, 18.70:1,18.75:1, 18.80:1, 18.85:1, 18.90:1, 18.95:1, 19.00:1, 19.05:1, 19.10:1,19.15:1, 19.20:1, 19.25:1, 19.30:1, 19.35:1, 19.40:1, 19.45:1, 19.50:1,19.55:1, 19.60:1, 19.65:1, 19.70:1, 19.75:1, 19.80:1, 19.85:1, 19.90:1,19.95:1, 20.00:1, 20.05:1, 20.10:1, 20.15:1, 20.20:1, 20.25:1, 20.30:1,20.35:1, 20.40:1, 20.45:1, 20.50:1, 20.55:1, 20.60:1, 20.65:1, 20.70:1,20.75:1, 20.80:1, 20.85:1, 20.90:1, 20.95:1, 21.00:1, 21.05:1, 21.10:1,21.15:1, 21.20:1, 21.25:1, 21.30:1, 21.35:1, 21.40:1, 21.45:1, 21.50:1,21.55:1, 21.60:1, 21.65:1, 21.70:1, 21.75:1, 21.80:1, 21.85:1, 21.90:1,21.95:1, 22.00:1, 22.05:1, 22.10:1, 22.15:1, 22.20:1, 22.25:1, 22.30:1,22.35:1, 22.40:1, 22.45:1, 22.50:1, 22.55:1, 22.60:1, 22.65:1, 22.70:1,22.75:1, 22.80:1, 22.85:1, 22.90:1, 22.95:1, 23.00, 23.05:1, 23.10:1,23.15:1, 23.20:1, 23.25:1, 23.30:1, 23.35:1, 23.40:1, 23.45:1, 23.50:1,23.55:1, 23.60:1, 23.65:1, 23.70:1, 23.75:1, 23.80:1, 23.85:1, 23.90:1,23.95:1, 24.00:1, 24.05:1, 24.10:1, 24.15:1, 24.20:1, 24.25:1, 24.30:1,24.35:1, 24.40:1, 24.45:1, 24.50:1, 24.55:1, 24.60:1, 24.65:1, 24.70:1,24.75:1, 24.80:1, 24.85:1, 24.90:1, 24.95:1, 25.05:1, 25.10:1, 25.15:1,25.20:1, 25.25:1, 25.30:1, 25.35:1, 25.40:1, 25.45:1, 25.50:1, 25.55:1,25.60:1, 25.65:1, 25.70:1, 25.75:1, 25.80:1, 25.85:1, 25.90:1, 25.95:1,26.00:1, 26.05:1, 26.10:1, 26.15:1, 26.20:1, 26.25:1, 26.30:1, 26.35:1,26.40:1, 26.45:1, 26.50:1, 26.55:1, 26.60:1, 26.65:1, 26.70:1, 26.75:1,26.80:1, 26.85:1, 26.90:1, or 26.95:1. In some embodiments, theperipheral blood of the donor is enriched with CD34⁺ CD90⁺ CD45RA⁻ cellsrelative to leukocytes by a ratio of about from about 5.5:1 to about6.5:1, such as a ratio of about 5.50:1, 5.55:1, 5.60:1, 5.65:1, 5.70:1,5.75:1, 5.80:1, 5.85:1, 5.90:1, 5.95:1, 6.00:1, 6.05:1, 6.10:1, 6.15:1,6.20:1, 6.25:1, 6.30:1, 6.35:1, 6.40:1, 6.45:1, or 6.50:1. In someembodiments, the peripheral blood of the donor is enriched with CD34⁺CD90⁺ CD45RA⁻ cells relative to leukocytes by a ratio of about 6.0:1.

In another aspect, the invention features a method of mobilizing apopulation of hematopoietic stem cells from the bone marrow of amammalian donor (e.g., a human donor) into peripheral blood, the methodincluding administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to produce a population of cells havinga density of CD34⁺ CD90⁺ CD45RA⁻ cells of at least about 16,000cells/ml, such as a density of from about 20,000 cells/ml to about75,000 cells/ml, about 25,000 cells/ml to about 70,000 cells/ml, about30,000 cells/ml to about 65,000 cells/ml, about 35,000 cells/ml to about60,000 cells/ml, about 40,000 cells/ml to about 55,000 cells/ml, orabout 45,000 cells/ml to about 50,000 cells/ml (e.g., about 16,000cells/ml, 17,000 cells/ml, 18,000 cells/ml, 19,000 cells/ml, 20,000cells/ml, 21,000 cells/ml, 22,000 cells/ml, 23,000 cells/ml, 24,000cells/ml, 25,000 cells/ml, 26,000 cells/ml, 27,000 cells/ml, 28,000cells/ml, 29,000 cells/ml, 30,000 cells/ml, 31,000 cells/ml, 32,000cells/ml, 33,000 cells/ml, 34,000 cells/ml, 35,000 cells/ml, 36,000cells/ml, 37,000 cells/ml, 38,000 cells/ml, 39,000 cells/ml, 40,000cells/ml, 41,000 cells/ml, 42,000 cells/ml, 43,000 cells/ml, 44,000cells/ml, 45,000 cells/ml, 46,000 cells/ml, 47,000 cells/ml, 48,000cells/ml, 49,000 cells/ml, 50,000 cells/ml, 51,000 cells/ml, 52,000cells/ml, 53,000 cells/ml, 54,000 cells/ml, 55,000 cells/ml, 56,000cells/ml, 57,000 cells/ml, 58,000 cells/ml, 59,000 cells/ml, 60,000cells/ml, 61,000 cells/ml, 62,000 cells/ml, 63,000 cells/ml, 64,000cells/ml, 65,000 cells/ml, 66,000 cells/ml, 67,000 cells/ml, 68,000cells/ml, 69,000 cells/ml, 70,000 cells/ml, 71,000 cells/ml, 72,000cells/ml, 73,000 cells/ml, 74,000 cells/ml, 75,000 cells/ml, 76,000cells/ml, 77,000 cells/ml, or more), and having a density of leukocytesof no more than about 5.3×10⁷ cells/ml, such as a density of leukocytesof about 2.3×10⁷ cells/ml to about 5.3×10⁷ cells/ml, about 2.5×10⁷cells/ml to about 5.1×10⁷ cells/ml, 2.9×10⁷ cells/ml to about 4.5×10⁷cells/ml, about 3×10⁷ cells/ml to about 4×10⁷ cells/ml (e.g., 5.3×10⁷cells/ml, 5.2×10⁷ cells/ml, 5.1×10⁷ cells/ml, 5×10⁷ cells/ml, 4.9×10⁷cells/ml, 4.8×10⁷ cells/ml, 4.7×10⁷ cells/ml, 4.6×10⁷ cells/ml, 4.5×10⁷cells/ml, 4.4×10⁷ cells/ml, 4.3×10⁷ cells/ml 4.2×10⁷ cells/ml, 4.1×10⁷cells/ml 4×10⁷ cells/ml, 3.9×10⁷ cells/ml, 3.8×10⁷ cells/ml, 3.7×10⁷cells/ml, 3.6×10⁷ cells/ml, 3.5×10⁷ cells/ml, 3.4×10⁷ cells/ml, 3.3×10⁷cells/ml, 3.2×10⁷ cells/ml, 3.1×10⁷ cells/ml, 3×10⁷ cells/ml, 2.9×10⁷cells/ml, 2.8×10⁷ cells/ml, 2.7×10⁷ cells/ml, 2.6×10⁷ cells/ml, 2.5×10⁷cells/ml, 2.4×10⁷ cells/ml, 2.3×10⁷ cells/ml, or less). In someembodiments, the method includes administering to the donor a CXCR2agonist and a CXCR4 antagonist in amounts sufficient to produce apopulation of cells having a density of CD34⁺ CD90⁺ CD45RA⁻ cells offrom about 20,000 cells/ml to about 75,000 cells/ml, and having adensity of leukocytes of from about 2.3×10⁷ cells/ml to about 5.3×10⁷cells/ml. In some embodiments, the method includes administering to thedonor a CXCR2 agonist and a CXCR4 antagonist in amounts sufficient toproduce a population of cells having a density of CD34⁺ CD90⁺ CD45RA⁻cells of from about 30,000 cells/ml to about 60,000 cells/ml, and havinga density of leukocytes of from about 2.5×10⁷ cells/ml to about 5×10⁷cells/ml. In some embodiments, the method includes administering to thedonor a CXCR2 agonist and a CXCR4 antagonist in amounts sufficient toproduce a population of cells having a density of CD34⁺ CD90⁺ CD45RA⁻cells of from about 40,000 cells/ml to about 50,000 cells/ml, and havinga density of leukocytes of from about 3×10⁷ cells/ml to about 4×10⁷cells/ml.

In a further aspect, the invention features a method of mobilizing apopulation of hematopoietic stem cells from the bone marrow of amammalian donor (e.g., a human donor) into peripheral blood, the methodincluding administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to produce a population of cells havinga ratio of CD34⁺ CD90⁺ CD45RA⁻ cells to neutrophils of from about 0.0007to about 0.0043 in a sample of peripheral blood of the donor followingadministration of the CXCR2 agonist and CXCR4 antagonist. In someembodiments, the ratio of CD34⁺ CD90⁺ CD45RA⁻ cells to neutrophils inthe sample may be about 0.00070, 0.00071, 0.00072, 0.00073, 0.00074,0.00075, 0.00076, 0.00077, 0.00078, 0.00079, 0.00080, 0.00081, 0.00082,0.00083, 0.00084, 0.00085, 0.00086, 0.00087, 0.00088, 0.00089, 0.00090,0.00091, 0.00092, 0.00093, 0.00094, 0.00095, 0.00096, 0.00097, 0.00098,0.00099, 0.00100, 0.00101, 0.00102, 0.00103, 0.00104, 0.00105, 0.00106,0.00107, 0.00108, 0.00109, 0.00110, 0.00111, 0.00112, 0.00113, 0.00114,0.00115, 0.00116, 0.00117, 0.00118, 0.00119, 0.00120, 0.00121, 0.00122,0.00123, 0.00124, 0.00125, 0.00126, 0.00127, 0.00128, 0.00129, 0.00130,0.00131, 0.00132, 0.00133, 0.00134, 0.00135, 0.00136, 0.00137, 0.00138,0.00139, 0.00140, 0.00141, 0.00142, 0.00143, 0.00144, 0.00145, 0.00146,0.00147, 0.00148, 0.00149, 0.00150, 0.00151, 0.00152, 0.00153, 0.00154,0.00155, 0.00156, 0.00157, 0.00158, 0.00159, 0.00160, 0.00161, 0.00162,0.00163, 0.00164, 0.00165, 0.00166, 0.00167, 0.00168, 0.00169, 0.00170,0.00171, 0.00172, 0.00173, 0.00174, 0.00175, 0.00176, 0.00177, 0.00178,0.00179, 0.00180, 0.00181, 0.00182, 0.00183, 0.00184, 0.00185, 0.00186,0.00187, 0.00188, 0.00189, 0.00190, 0.00191, 0.00192, 0.00193, 0.00194,0.00195, 0.00196, 0.00197, 0.00198, 0.00199, 0.00200, 0.00201, 0.00202,0.00203, 0.00204, 0.00205, 0.00206, 0.00207, 0.00208, 0.00209, 0.00210,0.00211, 0.00212, 0.00213, 0.00214, 0.00215, 0.00216, 0.00217, 0.00218,0.00219, 0.00220, 0.00221, 0.00222, 0.00223, 0.00224, 0.00225, 0.00226,0.00227, 0.00228, 0.00229, 0.00230, 0.00231, 0.00232, 0.00233, 0.00234,0.00235, 0.00236, 0.00237, 0.00238, 0.00239, 0.00240, 0.00241, 0.00242,0.00243, 0.00244, 0.00245, 0.00246, 0.00247, 0.00248, 0.00249, 0.00250,0.00251, 0.00252, 0.00253, 0.00254, 0.00255, 0.00256, 0.00257, 0.00258,0.00259, 0.00260, 0.00261, 0.00262, 0.00263, 0.00264, 0.00265, 0.00266,0.00267, 0.00268, 0.00269, 0.00270, 0.00271, 0.00272, 0.00273, 0.00274,0.00275, 0.00276, 0.00277, 0.00278, 0.00279, 0.00280, 0.00281, 0.00282,0.00283, 0.00284, 0.00285, 0.00286, 0.00287, 0.00288, 0.00289, 0.00290,0.00291, 0.00292, 0.00293, 0.00294, 0.00295, 0.00296, 0.00297, 0.00298,0.00299, 0.00300, 0.00300, 0.00301, 0.00302, 0.00303, 0.00304, 0.00305,0.00306, 0.00307, 0.00308, 0.00309, 0.00310, 0.00311, 0.00312, 0.00313,0.00314, 0.00315, 0.00316, 0.00317, 0.00318, 0.00319, 0.00320, 0.00321,0.00322, 0.00323, 0.00324, 0.00325, 0.00326, 0.00327, 0.00328, 0.00329,0.00330, 0.00331, 0.00332, 0.00333, 0.00334, 0.00335, 0.00336, 0.00337,0.00338, 0.00339, 0.00340, 0.00341, 0.00342, 0.00343, 0.00344, 0.00345,0.00346, 0.00347, 0.00348, 0.00349, 0.00350, 0.00351, 0.00352, 0.00353,0.00354, 0.00355, 0.00356, 0.00357, 0.00358, 0.00359, 0.00360, 0.00361,0.00362, 0.00363, 0.00364, 0.00365, 0.00366, 0.00367, 0.00368, 0.00369,0.00370, 0.00371, 0.00372, 0.00373, 0.00374, 0.00375, 0.00376, 0.00377,0.00378, 0.00379, 0.00380, 0.00381, 0.00382, 0.00383, 0.00384, 0.00385,0.00386, 0.00387, 0.00388, 0.00389, 0.00390, 0.00391, 0.00392, 0.00393,0.00394, 0.00395, 0.00396, 0.00397, 0.00398, 0.00399, 0.00400, 0.00401,0.00402, 0.00403, 0.00404, 0.00405, 0.00406, 0.00407, 0.00408, 0.00409,0.00410, 0.00411, 0.00412, 0.00413, 0.00414, 0.00415, 0.00416, 0.00417,0.00418, 0.00419, 0.00420, 0.00421, 0.00422, 0.00423, 0.00424, 0.00425,0.00426, 0.00427, 0.00428, 0.00429, or 0.00430. In some embodiments, theratio of CD34⁺ CD90⁺ CD45RA⁻ cells to neutrophils in the sample is fromabout 0.002 to about 0.003. In some embodiments, the ratio of CD34⁺CD90⁺ CD45RA⁻ cells to neutrophils in the sample is from about 0.0014 toabout 0.0034, such as a ratio of CD34⁺ CD90⁺ CD45RA⁻ cells toneutrophils in the sample of about 0.00140, 0.00141, 0.00142, 0.00143,0.00144, 0.00145, 0.00146, 0.00147, 0.00148, 0.00149, 0.00150, 0.00151,0.00152, 0.00153, 0.00154, 0.00155, 0.00156, 0.00157, 0.00158, 0.00159,0.00160, 0.00161, 0.00162, 0.00163, 0.00164, 0.00165, 0.00166, 0.00167,0.00168, 0.00169, 0.00170, 0.00171, 0.00172, 0.00173, 0.00174, 0.00175,0.00176, 0.00177, 0.00178, 0.00179, 0.00180, 0.00181, 0.00182, 0.00183,0.00184, 0.00185, 0.00186, 0.00187, 0.00188, 0.00189, 0.00190, 0.00191,0.00192, 0.00193, 0.00194, 0.00195, 0.00196, 0.00197, 0.00198, 0.00199,0.00200, 0.00201, 0.00202, 0.00203, 0.00204, 0.00205, 0.00206, 0.00207,0.00208, 0.00209, 0.00210, 0.00211, 0.00212, 0.00213, 0.00214, 0.00215,0.00216, 0.00217, 0.00218, 0.00219, 0.00220, 0.00221, 0.00222, 0.00223,0.00224, 0.00225, 0.00226, 0.00227, 0.00228, 0.00229, 0.00230, 0.00231,0.00232, 0.00233, 0.00234, 0.00235, 0.00236, 0.00237, 0.00238, 0.00239,0.00240, 0.00241, 0.00242, 0.00243, 0.00244, 0.00245, 0.00246, 0.00247,0.00248, 0.00249, 0.00250, 0.00251, 0.00252, 0.00253, 0.00254, 0.00255,0.00256, 0.00257, 0.00258, 0.00259, 0.00260, 0.00261, 0.00262, 0.00263,0.00264, 0.00265, 0.00266, 0.00267, 0.00268, 0.00269, 0.00270, 0.00271,0.00272, 0.00273, 0.00274, 0.00275, 0.00276, 0.00277, 0.00278, 0.00279,0.00280, 0.00281, 0.00282, 0.00283, 0.00284, 0.00285, 0.00286, 0.00287,0.00288, 0.00289, 0.00290, 0.00291, 0.00292, 0.00293, 0.00294, 0.00295,0.00296, 0.00297, 0.00298, 0.00299, 0.00300, 0.00300, 0.00301, 0.00302,0.00303, 0.00304, 0.00305, 0.00306, 0.00307, 0.00308, 0.00309, 0.00310,0.00311, 0.00312, 0.00313, 0.00314, 0.00315, 0.00316, 0.00317, 0.00318,0.00319, 0.00320, 0.00321, 0.00322, 0.00323, 0.00324, 0.00325, 0.00326,0.00327, 0.00328, 0.00329, 0.00330, 0.00331, 0.00332, 0.00333, 0.00334,0.00335, 0.00336, 0.00337, 0.00338, 0.00339, or 0.00340. In someembodiments, the ratio of CD34⁺ CD90⁺ CD45RA⁻ cells to neutrophils inthe sample is about 0.0024.

In an additional aspect, the invention features a method of mobilizing apopulation of hematopoietic stem cells from the bone marrow of amammalian donor (e.g., a human donor) into peripheral blood, the methodincluding administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to enrich the peripheral blood of thedonor with CD34⁺ CD90⁺ CD45RA⁻ cells relative to neutrophils by a ratioof from about 3.5:1 to about 22.0:1 as assessed by comparing a sample ofperipheral blood of the donor following administration of the CXCR2agonist and CXCR4 antagonist to a sample of peripheral blood of thedonor prior to administration of the CXCR2 agonist and CXCR4 antagonist.In some embodiments, the peripheral blood of the donor may be enrichedwith CD34⁺ CD90⁺ CD45RA⁻ cells relative to neutrophils by a ratio ofabout 3.50:1, 3.55:1, 3.60:1, 3.65:1, 3.70:1, 3.75:1, 3.80:1, 3.85:1,3.90:1, 3.95:1, 4.00:1, 4.05:1, 4.10:1, 4.15:1, 4.20:1, 4.25:1, 4.30:1,4.35:1, 4.40:1, 4.45:1, 4.50:1, 4.55:1, 4.60:1, 4.65:1, 4.70:1, 4.75:1,4.80:1, 4.85:1, 4.90:1, 4.95:1, 5.00:1, 5.05:1, 5.10:1, 5.15:1, 5.20:1,5.25:1, 5.30:1, 5.35:1, 5.40:1, 5.45:1, 5.50:1, 5.55:1, 5.60:1, 5.65:1,5.70:1, 5.75:1, 5.80:1, 5.85:1, 5.90:1, 5.95:1, 6.00:1, 6.05:1, 6.10:1,6.15:1, 6.20:1, 6.25:1, 6.30:1, 6.35:1, 6.40:1, 6.45:1, 6.50:1, 6.55:1,6.60:1, 6.65:1, 6.70:1, 6.75:1, 6.80:1, 6.85:1, 6.90:1, 6.95:1, 7.00:1,7.05:1, 7.10:1, 7.15:1, 7.20:1, 7.25:1, 7.30:1, 7.35:1, 7.40:1, 7.45:1,7.50:1, 7.55:1, 7.60:1, 7.65:1, 7.70:1, 7.75:1, 7.80:1, 7.85:1, 7.90:1,7.95:1, 8.00:1, 8.05:1, 8.10:1, 8.15:1, 8.20:1, 8.25:1, 8.30:1, 8.35:1,8.40:1, 8.45:1, 8.50:1, 8.55:1, 8.60:1, 8.65:1, 8.70:1, 8.75:1, 8.80:1,8.85:1, 8.90:1, 8.95:1, 9.00:1, 9.05:1, 9.10:1, 9.15:1, 9.20:1, 9.25:1,9.30:1, 9.35:1, 9.40:1, 9.45:1, 9.50:1, 9.55:1, 9.60:1, 9.65:1, 9.70:1,9.75:1, 9.80:1, 9.85:1, 9.90:1, 9.95:1, 10.00:1, 10.05:1, 10.10:1,10.15:1, 10.20:1, 10.25:1, 10.30:1, 10.35:1, 10.40:1, 10.45:1, 10.50:1,10.55:1, 10.60:1, 10.65:1, 10.70:1, 10.75:1, 10.80:1, 10.85:1, 10.90:1,10.95:1, 11.00:1, 11.05:1, 11.10:1, 11.15:1, 11.20:1, 11.25:1, 11.30:1,11.35:1, 11.40:1, 11.45:1, 11.50:1, 11.55:1, 11.60:1, 11.65:1, 11.70:1,11.75:1, 11.80:1, 11.85:1, 11.90:1, 11.95:1, 12.00:1, 12.05:1, 12.10:1,12.15:1, 12.20:1, 12.25:1, 12.30:1, 12.35:1, 12.40:1, 12.45:1, 12.50:1,12.55:1, 12.60:1, 12.65:1, 12.70:1, 12.75:1, 12.80:1, 12.85:1, 12.90:1,12.95:1, 13.00:1, 13.05:1, 13.10:1, 13.15:1, 13.20:1, 13.25:1, 13.30:1,13.35:1, 13.40:1, 13.45:1, 13.50:1, 13.55:1, 13.60:1, 13.65:1, 13.70:1,13.75:1, 13.80:1, 13.85:1, 13.90:1, 13.95:1, 14.00:1, 14.05:1, 14.10:1,14.15:1, 14.20:1, 14.25:1, 14.30:1, 14.35:1, 14.40:1, 14.45:1, 14.50:1,14.55:1, 14.60:1, 14.65:1, 14.70:1, 14.75:1, 14.80:1, 14.85:1, 14.90:1,14.95:1, 15.00:1, 15.05:1, 15.10:1, 15.15:1, 15.20:1, 15.25:1, 15.30:1,15.35:1, 15.40:1, 15.45:1, 15.50:1, 15.55:1, 15.60:1, 15.65:1, 15.70:1,15.75:1, 15.80:1, 15.85:1, 15.90:1, 15.95:1, 16.00:1, 16.05:1, 16.10:1,16.15:1, 16.20:1, 16.25:1, 16.30:1, 16.35:1, 16.40:1, 16.45:1, 16.50:1,16.55:1, 16.60:1, 16.65:1, 16.70:1, 16.75:1, 16.80:1, 16.85:1, 16.90:1,16.95:1, 17.00:1, 17.05:1, 17.10:1, 17.15:1, 17.20:1, 17.25:1, 17.30:1,17.35:1, 17.40:1, 17.45:1, 17.50:1, 17.55:1, 17.60:1, 17.65:1, 17.70:1,17.75:1, 17.80:1, 17.85:1, 17.90:1, 17.95:1, 18.00:1, 18.05:1, 18.10:1,18.15:1, 18.20:1, 18.25:1, 18.30:1, 18.35:1, 18.40:1, 18.45:1, 18.50:1,18.55:1, 18.60:1, 18.65:1, 18.70:1, 18.75:1, 18.80:1, 18.85:1, 18.90:1,18.95:1, 19.00:1, 19.05:1, 19.10:1, 19.15:1, 19.20:1, 19.25:1, 19.30:1,19.35:1, 19.40:1, 19.45:1, 19.50:1, 19.55:1, 19.60:1, 19.65:1, 19.70:1,19.75:1, 19.80:1, 19.85:1, 19.90:1, 19.95:1, 20.00:1, 20.05:1, 20.10:1,20.15:1, 20.20:1, 20.25:1, 20.30:1, 20.35:1, 20.40:1, 20.45:1, 20.50:1,20.55:1, 20.60:1, 20.65:1, 20.70:1, 20.75:1, 20.80:1, 20.85:1, 20.90:1,20.95:1, 21.00:1, 21.05:1, 21.10:1, 21.15:1, 21.20:1, 21.25:1, 21.30:1,21.35:1, 21.40:1, 21.45:1, 21.50:1, 21.55:1, 21.60:1, 21.65:1, 21.70:1,21.75:1, 21.80:1, 21.85:1, 21.90:1, 21.95:1, or 22.00:1. In someembodiments, the peripheral blood of the donor is enriched with CD34⁺CD90⁺ CD45RA⁻ cells relative to neutrophils by a ratio of about fromabout 7.0:1 to about 10:1. In some embodiments, the peripheral blood ofthe donor is enriched with CD34⁺ CD90⁺ CD45RA⁻ cells relative toneutrophils by a ratio of from about 7.00:1 to about 9.00:1, such as aratio of about 7.00:1, 7.05:1, 7.10:1, 7.15:1, 7.20:1, 7.25:1, 7.30:1,7.35:1, 7.40:1, 7.45:1, 7.50:1, 7.55:1, 7.60:1, 7.65:1, 7.70:1, 7.75:1,7.80:1, 7.85:1, 7.90:1, 7.95:1, 8.00:1, 8.05:1, 8.10:1, 8.15:1, 8.20:1,8.25:1, 8.30:1, 8.35:1, 8.40:1, 8.45:1, 8.50:1, 8.55:1, 8.60:1, 8.65:1,8.70:1, 8.75:1, 8.80:1, 8.85:1, 8.90:1, 8.95:1, or 9.00:1. In someembodiments, the peripheral blood of the donor is enriched with CD34⁺CD90⁺ CD45RA⁻ cells relative to neutrophils by a ratio of about 8.2:1.

In another aspect, the invention features a method of mobilizing apopulation of hematopoietic stem cells from the bone marrow of amammalian donor (e.g., a human donor) into peripheral blood, the methodincluding administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to produce a population of cells havinga density of CD34⁺ CD90⁺ CD45RA⁻ cells of at least about 16,000cells/ml, such as a density of from about 20,000 cells/ml to about75,000 cells/ml, about 25,000 cells/ml to about 70,000 cells/ml, about30,000 cells/ml to about 65,000 cells/ml, about 35,000 cells/ml to about60,000 cells/ml, about 40,000 cells/ml to about 55,000 cells/ml, orabout 45,000 cells/ml to about 50,000 cells/ml (e.g., about 16,000cells/ml, 17,000 cells/ml, 18,000 cells/ml, 19,000 cells/ml, 20,000cells/ml, 21,000 cells/ml, 22,000 cells/ml, 23,000 cells/ml, 24,000cells/ml, 25,000 cells/ml, 26,000 cells/ml, 27,000 cells/ml, 28,000cells/ml, 29,000 cells/ml, 30,000 cells/ml, 31,000 cells/ml, 32,000cells/ml, 33,000 cells/ml, 34,000 cells/ml, 35,000 cells/ml, 36,000cells/ml, 37,000 cells/ml, 38,000 cells/ml, 39,000 cells/ml, 40,000cells/ml, 41,000 cells/ml, 42,000 cells/ml, 43,000 cells/ml, 44,000cells/ml, 45,000 cells/ml, 46,000 cells/ml, 47,000 cells/ml, 48,000cells/ml, 49,000 cells/ml, 50,000 cells/ml, 51,000 cells/ml, 52,000cells/ml, 53,000 cells/ml, 54,000 cells/ml, 55,000 cells/ml, 56,000cells/ml, 57,000 cells/ml, 58,000 cells/ml, 59,000 cells/ml, 60,000cells/ml, 61,000 cells/ml, 62,000 cells/ml, 63,000 cells/ml, 64,000cells/ml, 65,000 cells/ml, 66,000 cells/ml, 67,000 cells/ml, 68,000cells/ml, 69,000 cells/ml, 70,000 cells/nil, 71,000 cells/ml, 72,000cells/ml, 73,000 cells/ml, 74,000 cells/ml, 75,000 cells/ml, 76,000cells/ml, 77,000 cells/ml, or more), and having a density of neutrophilsof no more than about 2.5×10⁷ cells/ml, such as a density of neutrophilsof about 1×10⁷ cells/ml to about 2.5×10⁷ cells/ml, about 1.3×10⁷cells/ml to about 2×10⁷ cells/ml, or about 1.5×10⁷ cells/ml to about1.9×10⁷ cells/ml (e.g., about 2.5×10⁷ cells/ml, 2.4×10⁷ cells/ml,2.3×10⁷ cells/ml, 2.2×10⁷ cells/ml, 2.1×10⁷ cells/ml, 2×10⁷ cells/ml,1.9×10⁷ cells/ml, 1.8×10⁷ cells/ml, 1.7×10⁷ cells/ml, 1.6×10⁷ cells/ml,1.5×10⁷ cells/ml 1.4×10⁷ cells/ml, 1.3×10⁷ cells/ml, 1.2×10⁷ cells/ml,1.1×10⁷ cells/ml, 1×10⁷ cells/ml, or less). In some embodiments, themethod includes administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to produce a population of cells havinga density of CD34⁺ CD90⁺ CD45RA⁻ cells of from about 20,000 cells/ml toabout 75,000 cells/ml, and having a density of neutrophils of from about1×10⁷ cells/ml to about 2.5×10⁷ cells/ml. In some embodiments, themethod includes administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to produce a population of cells havinga density of CD34⁺ CD90⁺ CD45RA⁻ cells of from about 30,000 cells/ml toabout 60,000 cells/ml, and having a density of neutrophils of from about1.3×10⁷ cells/ml to about 2.3×10⁷ cells/ml. In some embodiments, themethod includes administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to produce a population of cells havinga density of CD34⁺ CD90⁺ CD45RA⁻ cells of from about 40,000 cells/ml toabout 50,000 cells/ml, and having a density of neutrophils of from about1.5×10⁷ cells/ml to about 2×10⁷ cells/ml.

In yet another aspect, the invention features a method of mobilizing apopulation of hematopoietic stem cells from the bone marrow of amammalian donor (e.g., a human donor) into peripheral blood, the methodincluding administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to produce a population of cells havinga ratio of CD34⁺ CD90⁺ CD45RA⁻ cells to lymphocytes of from about 0.0008to about 0.0069 in a sample of peripheral blood of the donor followingadministration of the CXCR2 agonist and CXCR4 antagonist. In someembodiments, the ratio of CD34⁺ CD90⁺ CD45RA⁻ cells to lymphocytes inthe sample may be about 0.00080, 0.00081, 0.00082, 0.00083, 0.00084,0.00085, 0.00086, 0.00087, 0.00088, 0.00089, 0.00090, 0.00091, 0.00092,0.00093, 0.00094, 0.00095, 0.00096, 0.00097, 0.00098, 0.00099, 0.00100,0.00101, 0.00102, 0.00103, 0.00104, 0.00105, 0.00106, 0.00107, 0.00108,0.00109, 0.00110, 0.00111, 0.00112, 0.00113, 0.00114, 0.00115, 0.00116,0.00117, 0.00118, 0.00119, 0.00120, 0.00121, 0.00122, 0.00123, 0.00124,0.00125, 0.00126, 0.00127, 0.00128, 0.00129, 0.00130, 0.00131, 0.00132,0.00133, 0.00134, 0.00135, 0.00136, 0.00137, 0.00138, 0.00139, 0.00140,0.00141, 0.00142, 0.00143, 0.00144, 0.00145, 0.00146, 0.00147, 0.00148,0.00149, 0.00150, 0.00151, 0.00152, 0.00153, 0.00154, 0.00155, 0.00156,0.00157, 0.00158, 0.00159, 0.00160, 0.00161, 0.00162, 0.00163, 0.00164,0.00165, 0.00166, 0.00167, 0.00168, 0.00169, 0.00170, 0.00171, 0.00172,0.00173, 0.00174, 0.00175, 0.00176, 0.00178, 0.00179, 0.00180, 0.00181,0.00182, 0.00183, 0.00184, 0.00185, 0.00186, 0.00187, 0.00188, 0.00189,0.00190, 0.00191, 0.00192, 0.00193, 0.00194, 0.00195, 0.00196, 0.00197,0.00198, 0.00199, 0.00200, 0.00201, 0.00202, 0.00203, 0.00204, 0.00205,0.00206, 0.00207, 0.00208, 0.00209, 0.00210, 0.00211, 0.00212, 0.00213,0.00214, 0.00215, 0.00216, 0.00217, 0.00218, 0.00219, 0.00220, 0.00221,0.00222, 0.00223, 0.00224, 0.00225, 0.00226, 0.00227, 0.00228, 0.00229,0.00230, 0.00231, 0.00232, 0.00233, 0.00234, 0.00235, 0.00236, 0.00237,0.00238, 0.00239, 0.00240, 0.00241, 0.00242, 0.00243, 0.00244, 0.00245,0.00246, 0.00247, 0.00248, 0.00249, 0.00250, 0.00251, 0.00252, 0.00253,0.00254, 0.00255, 0.00256, 0.00257, 0.00258, 0.00259, 0.00260, 0.00261,0.00262, 0.00263, 0.00264, 0.00265, 0.00266, 0.00267, 0.00268, 0.00269,0.00270, 0.00271, 0.00272, 0.00273, 0.00274, 0.00275, 0.00276, 0.00278,0.00279, 0.00280, 0.00281, 0.00282, 0.00283, 0.00284, 0.00285, 0.00286,0.00287, 0.00288, 0.00289, 0.00290, 0.00291, 0.00292, 0.00293, 0.00294,0.00295, 0.00296, 0.00297, 0.00298, 0.00299, 0.00300, 0.00301, 0.00302,0.00303, 0.00304, 0.00305, 0.00306, 0.00307, 0.00308, 0.00309, 0.00310,0.00311, 0.00312, 0.00313, 0.00314, 0.00315, 0.00316, 0.00317, 0.00318,0.00319, 0.00320, 0.00321, 0.00322, 0.00323, 0.00324, 0.00325, 0.00326,0.00327, 0.00328, 0.00329, 0.00330, 0.00331, 0.00332, 0.00333, 0.00334,0.00335, 0.00336, 0.00337, 0.00338, 0.00339, 0.00340, 0.00341, 0.00342,0.00343, 0.00344, 0.00345, 0.00346, 0.00347, 0.00348, 0.00349, 0.00350,0.00351, 0.00352, 0.00353, 0.00354, 0.00355, 0.00356, 0.00357, 0.00358,0.00359, 0.00360, 0.00361, 0.00362, 0.00363, 0.00364, 0.00365, 0.00366,0.00367, 0.00368, 0.00369, 0.00370, 0.00371, 0.00372, 0.00373, 0.00374,0.00375, 0.00376, 0.00378, 0.00379, 0.00380, 0.00381, 0.00382, 0.00383,0.00384, 0.00385, 0.00386, 0.00387, 0.00388, 0.00389, 0.00390, 0.00391,0.00392, 0.00393, 0.00394, 0.00395, 0.00396, 0.00397, 0.00398, 0.00399,0.00401, 0.00402, 0.00403, 0.00404, 0.00405, 0.00406, 0.00407, 0.00408,0.00409, 0.00410, 0.00411, 0.00412, 0.00413, 0.00414, 0.00415, 0.00416,0.00417, 0.00418, 0.00419, 0.00420, 0.00421, 0.00422, 0.00423, 0.00424,0.00425, 0.00426, 0.00427, 0.00428, 0.00429, 0.00430, 0.00431, 0.00432,0.00433, 0.00434, 0.00435, 0.00436, 0.00437, 0.00438, 0.00439, 0.00440,0.00441, 0.00442, 0.00443, 0.00444, 0.00445, 0.00446, 0.00447, 0.00448,0.00449, 0.00450, 0.00451, 0.00452, 0.00453, 0.00454, 0.00455, 0.00456,0.00457, 0.00458, 0.00459, 0.00460, 0.00461, 0.00462, 0.00463, 0.00464,0.00465, 0.00466, 0.00467, 0.00468, 0.00469, 0.00470, 0.00471, 0.00472,0.00473, 0.00474, 0.00475, 0.00476, 0.00478, 0.00479, 0.00480, 0.00481,0.00482, 0.00483, 0.00484, 0.00485, 0.00486, 0.00487, 0.00488, 0.00489,0.00490, 0.00491, 0.00492, 0.00493, 0.00494, 0.00495, 0.00496, 0.00497,0.00498, 0.00499, 0.00500, 0.00501, 0.00502, 0.00503, 0.00504, 0.00505,0.00506, 0.00507, 0.00508, 0.00509, 0.00510, 0.00511, 0.00512, 0.00513,0.00514, 0.00515, 0.00516, 0.00517, 0.00518, 0.00519, 0.00520, 0.00521,0.00522, 0.00523, 0.00524, 0.00525, 0.00526, 0.00527, 0.00528, 0.00529,0.00530, 0.00531, 0.00532, 0.00533, 0.00534, 0.00535, 0.00536, 0.00537,0.00538, 0.00539, 0.00540, 0.00541, 0.00542, 0.00543, 0.00544, 0.00545,0.00546, 0.00547, 0.00548, 0.00549, 0.00550, 0.00551, 0.00552, 0.00553,0.00554, 0.00555, 0.00556, 0.00557, 0.00558, 0.00559, 0.00560, 0.00561,0.00562, 0.00563, 0.00564, 0.00565, 0.00566, 0.00567, 0.00568, 0.00569,0.00570, 0.00571, 0.00572, 0.00573, 0.00574, 0.00575, 0.00576, 0.00578,0.00579, 0.00580, 0.00581, 0.00582, 0.00583, 0.00584, 0.00585, 0.00586,0.00587, 0.00588, 0.00589, 0.00590, 0.00591, 0.00592, 0.00593, 0.00594,0.00595, 0.00596, 0.00597, 0.00598, 0.00599, 0.00600, 0.00601, 0.00602,0.00603, 0.00604, 0.00605, 0.00606, 0.00607, 0.00608, 0.00609, 0.00610,0.00611, 0.00612, 0.00613, 0.00614, 0.00615, 0.00616, 0.00617, 0.00618,0.00619, 0.00620, 0.00621, 0.00622, 0.00623, 0.00624, 0.00625, 0.00626,0.00627, 0.00628, 0.00629, 0.00630, 0.00631, 0.00632, 0.00633, 0.00634,0.00635, 0.00636, 0.00637, 0.00638, 0.00639, 0.00640, 0.00641, 0.00642,0.00643, 0.00644, 0.00645, 0.00646, 0.00647, 0.00648, 0.00649, 0.00650,0.00651, 0.00652, 0.00653, 0.00654, 0.00655, 0.00656, 0.00657, 0.00658,0.00659, 0.00660, 0.00661, 0.00662, 0.00663, 0.00664, 0.00665, 0.00666,0.00667, 0.00668, 0.00669, 0.00670, 0.00671, 0.00672, 0.00673, 0.00674,0.00675, 0.00676, 0.00678, 0.00679, 0.00680, 0.00681, 0.00682, 0.00683,0.00684, 0.00685, 0.00686, 0.00687, 0.00688, 0.00689, or 0.00690. Insome embodiments, the ratio of CD34⁺ CD90⁺ CD45RA⁻ cells to lymphocytesin the sample is from about 0.0011 to about 0.0031, such as a ratio ofCD34⁺ CD90⁺ CD45RA⁻ cells to lymphocytes in the sample of about 0.00110,0.00111, 0.00112, 0.00113, 0.00114, 0.00115, 0.00116, 0.00117, 0.00118,0.00119, 0.00120, 0.00121, 0.00122, 0.00123, 0.00124, 0.00125, 0.00126,0.00127, 0.00128, 0.00129, 0.00130, 0.00131, 0.00132, 0.00133, 0.00134,0.00135, 0.00136, 0.00137, 0.00138, 0.00139, 0.00140, 0.00141, 0.00142,0.00143, 0.00144, 0.00145, 0.00146, 0.00147, 0.00148, 0.00149, 0.00150,0.00151, 0.00152, 0.00153, 0.00154, 0.00155, 0.00156, 0.00157, 0.00158,0.00159, 0.00160, 0.00161, 0.00162, 0.00163, 0.00164, 0.00165, 0.00166,0.00167, 0.00168, 0.00169, 0.00170, 0.00171, 0.00172, 0.00173, 0.00174,0.00175, 0.00176, 0.00178, 0.00179, 0.00180, 0.00181, 0.00182, 0.00183,0.00184, 0.00185, 0.00186, 0.00187, 0.00188, 0.00189, 0.00190, 0.00191,0.00192, 0.00193, 0.00194, 0.00195, 0.00196, 0.00197, 0.00198, 0.00199,0.00200, 0.00201, 0.00202, 0.00203, 0.00204, 0.00205, 0.00206, 0.00207,0.00208, 0.00209, 0.00210, 0.00211, 0.00212, 0.00213, 0.00214, 0.00215,0.00216, 0.00217, 0.00218, 0.00219, 0.00220, 0.00221, 0.00222, 0.00223,0.00224, 0.00225, 0.00226, 0.00227, 0.00228, 0.00229, 0.00230, 0.00231,0.00232, 0.00233, 0.00234, 0.00235, 0.00236, 0.00237, 0.00238, 0.00239,0.00240, 0.00241, 0.00242, 0.00243, 0.00244, 0.00245, 0.00246, 0.00247,0.00248, 0.00249, 0.00250, 0.00251, 0.00252, 0.00253, 0.00254, 0.00255,0.00256, 0.00257, 0.00258, 0.00259, 0.00260, 0.00261, 0.00262, 0.00263,0.00264, 0.00265, 0.00266, 0.00267, 0.00268, 0.00269, 0.00270, 0.00271,0.00272, 0.00273, 0.00274, 0.00275, 0.00276, 0.00278, 0.00279, 0.00280,0.00281, 0.00282, 0.00283, 0.00284, 0.00285, 0.00286, 0.00287, 0.00288,0.00289, 0.00290, 0.00291, 0.00292, 0.00293, 0.00294, 0.00295, 0.00296,0.00297, 0.00298, 0.00299, 0.00300, 0.00301, 0.00302, 0.00303, 0.00304,0.00305, 0.00306, 0.00307, 0.00308, 0.00309, or 0.00310. In someembodiments, the ratio of CD34⁺ CD90⁺ CD45RA⁻ cells to lymphocytes inthe sample is about 0.0021.

In an additional aspect, the invention features a method of mobilizing apopulation of hematopoietic stem cells from the bone marrow of amammalian donor (e.g., a human donor) into peripheral blood, the methodincluding administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to enrich the peripheral blood of thedonor with CD34⁺ CD90⁺ CD45RA⁻ cells relative to lymphocytes by a ratioof from about 5.6:1 to about 37.0:1 as assessed by comparing a sample ofperipheral blood of the donor following administration of the CXCR2agonist and CXCR4 antagonist to a sample of peripheral blood of thedonor prior to administration of the CXCR2 agonist and CXCR4 antagonist.In some embodiments, the peripheral blood of the donor may be enrichedwith CD34⁺ CD90⁺ CD45RA⁻ cells relative to lymphocytes by a ratio ofabout 5.60:1, 5.65:1, 5.70:1, 5.75:1, 5.80:1, 5.85:1, 5.90:1, 5.95:1,6.00:1, 6.05:1, 6.10:1, 6.15:1, 6.20:1, 6.25:1, 6.30:1, 6.35:1, 6.40:1,6.45:1, 6.50:1, 6.55:1, 6.60:1, 6.65:1, 6.70:1, 6.75:1, 6.80:1, 6.85:1,6.90:1, 6.95:1, 7.00:1, 7.05:1, 7.10:1, 7.15:1, 7.20:1, 7.25:1, 7.30:1,7.35:1, 7.40:1, 7.45:1, 7.50:1, 7.55:1, 7.60:1, 7.65:1, 7.70:1, 7.75:1,7.80:1, 7.85:1, 7.90:1, 7.95:1, 8.00:1, 8.05:1, 8.10:1, 8.15:1, 8.20:1,8.25:1, 8.30:1, 8.35:1, 8.40:1, 8.45:1, 8.50:1, 8.55:1, 8.60:1, 8.65:1,8.70:1, 8.75:1, 8.80:1, 8.85:1, 8.90:1, 8.95:1, 9.00:1, 9.05:1, 9.10:1,9.15:1, 9.20:1, 9.25:1, 9.30:1, 9.35:1, 9.40:1, 9.45:1, 9.50:1, 9.55:1,9.60:1, 9.65:1, 9.70:1, 9.75:1, 9.80:1, 9.85:1, 9.90:1, 9.95:1, 10.00:1,10.05:1, 10.10:1, 10.15:1, 10.20:1, 10.25:1, 10.30:1, 10.35:1, 10.40:1,10.45:1, 10.50:1, 10.55:1, 10.60:1, 10.65:1, 10.70:1, 10.75:1, 10.80:1,10.85:1, 10.90:1, 10.95:1, 11.00:1, 11.05:1, 11.10:1, 11.15:1, 11.20:1,11.25:1, 11.30:1, 11.35:1, 11.40:1, 11.45:1, 11.50:1, 11.55:1, 11.60:1,11.65:1, 11.70:1, 11.75:1, 11.80:1, 11.85:1, 11.90:1, 11.95:1, 12.00:1,12.05:1, 12.10:1, 12.15:1, 12.20:1, 12.25:1, 12.30:1, 12.35:1, 12.40:1,12.45:1, 12.50:1, 12.55:1, 12.60:1, 12.65:1, 12.70:1, 12.75:1, 12.80:1,12.85:1, 12.90:1, 12.95:1, 13.00:1, 13.05:1, 13.10:1, 13.15:1, 13.20:1,13.25:1, 13.30:1, 13.35:1, 13.40:1, 13.45:1, 13.50:1, 13.55:1, 13.60:1,13.65:1, 13.70:1, 13.75:1, 13.80:1, 13.85:1, 13.90:1, 13.95:1, 14.00:1,14.05:1, 14.10:1, 14.15:1, 14.20:1, 14.25:1, 14.30:1, 14.35:1, 14.40:1,14.45:1, 14.50:1, 14.55:1, 14.60:1, 14.65:1, 14.70:1, 14.75:1, 14.80:1,14.85:1, 14.90:1, 14.95:1, 15.00:1, 15.05:1, 15.10:1, 15.15:1, 15.20:1,15.25:1, 15.30:1, 15.35:1, 15.40:1, 15.45:1, 15.50:1, 15.55:1, 15.60:1,15.65:1, 15.70:1, 15.75:1, 15.80:1, 15.85:1, 15.90:1, 15.95:1, 16.00:1,16.05:1, 16.10:1, 16.15:1, 16.20:1, 16.25:1, 16.30:1, 16.35:1, 16.40:1,16.45:1, 16.50:1, 16.55:1, 16.60:1, 16.65:1, 16.70:1, 16.75:1, 16.80:1,16.85:1, 16.90:1, 16.95:1, 17.00:1, 17.05:1, 17.10:1, 17.15:1, 17.20:1,17.25:1, 17.30:1, 17.35:1, 17.40:1, 17.45:1, 17.50:1, 17.55:1, 17.60:1,17.65:1, 17.70:1, 17.75:1, 17.80:1, 17.85:1, 17.90:1, 17.95:1, 18.00:1,18.05:1, 18.10:1, 18.15:1, 18.20:1, 18.25:1, 18.30:1, 18.35:1, 18.40:1,18.45:1, 18.50:1, 18.55:1, 18.60:1, 18.65:1, 18.70:1, 18.75:1, 18.80:1,18.85:1, 18.90:1, 18.95:1, 19.00:1, 19.05:1, 19.10:1, 19.15:1, 19.20:1,19.25:1, 19.30:1, 19.35:1, 19.40:1, 19.45:1, 19.50:1, 19.55:1, 19.60:1,19.65:1, 19.70:1, 19.75:1, 19.80:1, 19.85:1, 19.90:1, 19.95:1, 20.00:1,20.05:1, 20.10:1, 20.15:1, 20.20:1, 20.25:1, 20.30:1, 20.35:1, 20.40:1,20.45:1, 20.50:1, 20.55:1, 20.60:1, 20.65:1, 20.70:1, 20.75:1, 20.80:1,20.85:1, 20.90:1, 20.95:1, 21.00:1, 21.05:1, 21.10:1, 21.15:1, 21.20:1,21.25:1, 21.30:1, 21.35:1, 21.40:1, 21.45:1, 21.50:1, 21.55:1, 21.60:1,21.65:1, 21.70:1, 21.75:1, 21.80:1, 21.85:1, 21.90:1, 21.95:1, 22.00:1,22.05:1, 22.10:1, 22.15:1, 22.20:1, 22.25:1, 22.30:1, 22.35:1, 22.40:1,22.45:1, 22.50:1, 22.55:1, 22.60:1, 22.65:1, 22.70:1, 22.75:1, 22.80:1,22.85:1, 22.90:1, 22.95:1, 23.00, 23.05:1, 23.10:1, 23.15:1, 23.20:1,23.25:1, 23.30:1, 23.35:1, 23.40:1, 23.45:1, 23.50:1, 23.55:1, 23.60:1,23.65:1, 23.70:1, 23.75:1, 23.80:1, 23.85:1, 23.90:1, 23.95:1, 24.00:1,24.05:1, 24.10:1, 24.15:1, 24.20:1, 24.25:1, 24.30:1, 24.35:1, 24.40:1,24.45:1, 24.50:1, 24.55:1, 24.60:1, 24.65:1, 24.70:1, 24.75:1, 24.80:1,24.85:1, 24.90:1, 24.95:1, 25.05:1, 25.10:1, 25.15:1, 25.20:1, 25.25:1,25.30:1, 25.35:1, 25.40:1, 25.45:1, 25.50:1, 25.55:1, 25.60:1, 25.65:1,25.70:1, 25.75:1, 25.80:1, 25.85:1, 25.90:1, 25.95:1, 26.00:1, 26.05:1,26.10:1, 26.15:1, 26.20:1, 26.25:1, 26.30:1, 26.35:1, 26.40:1, 26.45:1,26.50:1, 26.55:1, 26.60:1, 26.65:1, 26.70:1, 26.75:1, 26.80:1, 26.85:1,26.90:1, 26.95:1, 27.00:1, 27.05:1, 27.10:1, 27.15:1, 27.20:1, 27.25:1,27.30:1, 27.35:1, 27.40:1, 27.45:1, 27.50:1, 27.55:1, 27.60:1, 27.65:1,27.70:1, 27.75:1, 27.80:1, 27.85:1, 27.90:1, 27.95:1, 28.00:1, 28.05:1,28.10:1, 28.15:1, 28.20:1, 28.25:1, 28.30:1, 28.35:1, 28.40:1, 28.45:1,28.50:1, 28.55:1, 28.60:1, 28.65:1, 28.70:1, 28.75:1, 28.80:1, 28.85:1,28.90:1, 28.95:1, 29.00:1, 29.05:1, 29.10:1, 29.15:1, 29.20:1, 29.25:1,29.30:1, 29.35:1, 29.40:1, 29.45:1, 29.50:1, 29.55:1, 29.60:1, 29.65:1,29.70:1, 29.75:1, 29.80:1, 29.85:1, 29.90:1, 29.95:1, 30.00:1, 30.05:1,30.10:1, 30.15:1, 30.20:1, 30.25:1, 30.30:1, 30.35:1, 30.40:1, 30.45:1,30.50:1, 30.55:1, 30.60:1, 30.65:1, 30.70:1, 30.75:1, 30.80:1, 30.85:1,30.90:1, 30.95:1, 31.00:1, 31.05:1, 31.10:1, 31.15:1, 31.20:1, 31.25:1,31.30:1, 31.35:1, 31.40:1, 31.45:1, 31.50:1, 31.55:1, 31.60:1, 31.65:1,31.70:1, 31.75:1, 31.80:1, 31.85:1, 31.90:1, 31.95:1, 32.00:1, 32.05:1,32.10:1, 32.15:1, 32.20:1, 32.25:1, 32.30:1, 32.35:1, 32.40:1, 32.45:1,32.50:1, 32.55:1, 32.60:1, 32.65:1, 32.70:1, 32.75:1, 32.80:1, 32.85:1,32.90:1, 32.95:1, 33.00:1, 33.05:1, 33.10:1, 33.15:1, 33.20:1, 33.25:1,33.30:1, 33.35:1, 33.40:1, 33.45:1, 33.50:1, 33.55:1, 33.60:1, 33.65:1,33.70:1, 33.75:1, 33.80:1, 33.85:1, 33.90:1, 33.95:1, 34.00:1, 34.05:1,34.10:1, 34.15:1, 34.20:1, 34.25:1, 34.30:1, 34.35:1, 34.40:1, 34.45:1,34.50:1, 34.55:1, 34.60:1, 34.65:1, 34.70:1, 34.75:1, 34.80:1, 34.85:1,34.90:1, 34.95:1, 35.00:1, 35.05:1, 35.10:1, 35.15:1, 35.20:1, 35.25:1,35.30:1, 35.35:1, 35.40:1, 35.45:1, 35.50:1, 35.55:1, 35.60:1, 35.65:1,35.70:1, 35.75:1, 35.80:1, 35.85:1, 35.90:1, 35.95:1, 36.00:1, 36.05:1,36.10:1, 36.15:1, 36.20:1, 36.25:1, 36.30:1, 36.35:1, 36.40:1, 36.45:1,36.50:1, 36.55:1, 36.60:1, 36.65:1, 36.70:1, 36.75:1, 36.80:1, 36.85:1,36.90:1, 36.95:1, or 37.00. In some embodiments, the peripheral blood ofthe donor is enriched with CD34⁺ CD90⁺ CD45RA⁻ cells relative tolymphocytes by a ratio of about from about 8.0:1 to about 10.0:1, suchas a ratio of about 8.00:1, 8.05:1, 8.10:1, 8.15:1, 8.20:1, 8.25:1,8.30:1, 8.35:1, 8.40:1, 8.45:1, 8.50:1, 8.55:1, 8.60:1, 8.65:1, 8.70:1,8.75:1, 8.80:1, 8.85:1, 8.90:1, 8.95:1, 9.00:1, 9.05:1, 9.10:1, 9.15:1,9.20:1, 9.25:1, 9.30:1, 9.35:1, 9.40:1, 9.45:1, 9.50:1, 9.55:1, 9.60:1,9.65:1, 9.70:1, 9.75:1, 9.80:1, 9.85:1, 9.90:1, 9.95:1, or 10.00:1. Insome embodiments, the peripheral blood of the donor is enriched withCD34⁺ CD90⁺ CD45RA⁻ cells relative to lymphocytes by a ratio of about9.3:1.

In another aspect, the invention features a method of mobilizing apopulation of hematopoietic stem cells from the bone marrow of amammalian donor (e.g., a human donor) into peripheral blood, the methodincluding administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to produce a population of cells havinga density of CD34⁺ CD90⁺ CD45RA⁻ cells of at least about 16,000cells/ml, such as a density of from about 20,000 cells/ml to about75,000 cells/ml, about 25,000 cells/ml to about 70,000 cells/ml, about30,000 cells/ml to about 65,000 cells/ml, about 35,000 cells/ml to about60,000 cells/ml, about 40,000 cells/ml to about 55,000 cells/ml, orabout 45,000 cells/ml to about 50,000 cells/ml (e.g., about 16,000cells/ml, 17,000 cells/ml, 18,000 cells/ml, 19,000 cells/ml, 20,000cells/ml, 21,000 cells/ml, 22,000 cells/ml, 23,000 cells/ml, 24,000cells/ml, 25,000 cells/ml, 26,000 cells/ml, 27,000 cells/ml, 28,000cells/ml, 29,000 cells/ml, 30,000 cells/ml, 31,000 cells/ml, 32,000cells/ml, 33,000 cells/ml, 34,000 cells/ml, 35,000 cells/ml, 36,000cells/ml, 37,000 cells/ml, 38,000 cells/ml, 39,000 cells/ml, 40,000cells/ml, 41,000 cells/ml, 42,000 cells/ml, 43,000 cells/ml, 44,000cells/ml, 45,000 cells/ml, 46,000 cells/ml, 47,000 cells/ml, 48,000cells/ml, 49,000 cells/ml, 50,000 cells/ml, 51,000 cells/ml, 52,000cells/ml, 53,000 cells/ml, 54,000 cells/ml, 55,000 cells/ml, 56,000cells/ml, 57,000 cells/ml, 58,000 cells/ml, 59,000 cells/ml, 60,000cells/ml, 61,000 cells/ml, 62,000 cells/ml, 63,000 cells/ml, 64,000cells/ml, 65,000 cells/ml, 66,000 cells/ml, 67,000 cells/ml, 68,000cells/ml, 69,000 cells/ml, 70,000 cells/ml, 71,000 cells/ml, 72,000cells/ml, 73,000 cells/ml, 74,000 cells/ml, 75,000 cells/ml, 76,000cells/ml, 77,000 cells/ml, or more), and having a density of lymphocytesof no more than about 2.4×10⁷ cells/ml, such as a density of lymphocytesof about 1×10⁷ cells/ml to about 2.3×10⁷ cells/ml, about 1.3×10⁷cells/ml to about 2.1×10⁷ cells/ml, or about 1.5×10⁷ cells/ml to about1.9×10⁷ cells/ml (e.g., about 2.4×10⁷ cells/ml, 2.3×10⁷ cells/ml,2.2×10⁷ cells/ml, 2.1×10⁷ cells/ml, 2×10⁷ cells/ml, 1.9×10⁷ cells/ml,1.8×10⁷ cells/ml, 1.7×10⁷ cells/ml, 1.6×10⁷ cells/ml, 1.5×10⁷ cells/ml1.4×10⁷ cells/ml, 1.3×10⁷ cells/ml, 1.2×10⁷ cells/ml, 1.1×10⁷ cells/ml,1×10⁷ cells/ml, or less, 0.9×10⁷ cells/ml, 0.8×10⁷ cells/ml, or less).In some embodiments, the method includes administering to the donor aCXCR2 agonist and a CXCR4 antagonist in amounts sufficient to produce apopulation of cells having a density of CD34⁺ CD90⁺ CD45RA⁻ cells offrom about 20,000 cells/ml to about 75,000 cells/ml, and having adensity of lymphocytes of from about 1×10⁷ cells/ml to about 2.3×10⁷cells/ml. In some embodiments, the method includes administering to thedonor a CXCR2 agonist and a CXCR4 antagonist in amounts sufficient toproduce a population of cells having a density of CD34⁺ CD90⁺ CD45RA⁻cells of from about 30,000 cells/ml to about 60,000 cells/ml, and havinga density of lymphocytes of from about 1.3×10⁷ cells/ml to about 2.3×10⁷cells/ml. In some embodiments, the method includes administering to thedonor a CXCR2 agonist and a CXCR4 antagonist in amounts sufficient toproduce a population of cells having a density of CD34⁺ CD90⁺ CD45RA⁻cells of from about 40,000 cells/ml to about 50,000 cells/ml, and havinga density of lymphocytes of from about 1.5×10⁷ cells/ml to about 2×10⁷cells/ml.

In another aspect, the invention features a method of mobilizing apopulation of hematopoietic stem cells from the bone marrow of amammalian donor (e.g., a human donor) into peripheral blood, the methodincluding administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to produce a population of cells havinga ratio of CD34⁺ CD90⁺ CD45RA⁻ cells to monocytes of from about 0.0028to about 0.0130 in a sample of peripheral blood of the donor followingadministration of the CXCR2 agonist. In some embodiments, the ratio ofCD34⁺ CD90⁺ CD45RA⁻ cells to monocytes in the sample may be about0.00280, 0.00281, 0.00282, 0.00283, 0.00284, 0.00285, 0.00286, 0.00287,0.00288, 0.00289, 0.00290, 0.00291, 0.00292, 0.00293, 0.00294, 0.00295,0.00296, 0.00297, 0.00298, 0.00299, 0.00300, 0.00301, 0.00302, 0.00303,0.00304, 0.00305, 0.00306, 0.00307, 0.00308, 0.00309, 0.00310, 0.00311,0.00312, 0.00313, 0.00314, 0.00315, 0.00316, 0.00317, 0.00318, 0.00319,0.00320, 0.00321, 0.00322, 0.00323, 0.00324, 0.00325, 0.00326, 0.00327,0.00328, 0.00329, 0.00330, 0.00331, 0.00332, 0.00333, 0.00334, 0.00335,0.00336, 0.00337, 0.00338, 0.00339, 0.00340, 0.00341, 0.00342, 0.00343,0.00344, 0.00345, 0.00346, 0.00347, 0.00348, 0.00349, 0.00350, 0.00351,0.00352, 0.00353, 0.00354, 0.00355, 0.00356, 0.00357, 0.00358, 0.00359,0.00360, 0.00361, 0.00362, 0.00363, 0.00364, 0.00365, 0.00366, 0.00367,0.00368, 0.00369, 0.00370, 0.00371, 0.00372, 0.00373, 0.00374, 0.00375,0.00376, 0.00378, 0.00379, 0.00380, 0.00381, 0.00382, 0.00383, 0.00384,0.00385, 0.00386, 0.00387, 0.00388, 0.00389, 0.00390, 0.00391, 0.00392,0.00393, 0.00394, 0.00395, 0.00396, 0.00397, 0.00398, 0.00399, 0.00401,0.00402, 0.00403, 0.00404, 0.00405, 0.00406, 0.00407, 0.00408, 0.00409,0.00410, 0.00411, 0.00412, 0.00413, 0.00414, 0.00415, 0.00416, 0.00417,0.00418, 0.00419, 0.00420, 0.00421, 0.00422, 0.00423, 0.00424, 0.00425,0.00426, 0.00427, 0.00428, 0.00429, 0.00430, 0.00431, 0.00432, 0.00433,0.00434, 0.00435, 0.00436, 0.00437, 0.00438, 0.00439, 0.00440, 0.00441,0.00442, 0.00443, 0.00444, 0.00445, 0.00446, 0.00447, 0.00448, 0.00449,0.00450, 0.00451, 0.00452, 0.00453, 0.00454, 0.00455, 0.00456, 0.00457,0.00458, 0.00459, 0.00460, 0.00461, 0.00462, 0.00463, 0.00464, 0.00465,0.00466, 0.00467, 0.00468, 0.00469, 0.00470, 0.00471, 0.00472, 0.00473,0.00474, 0.00475, 0.00476, 0.00478, 0.00479, 0.00480, 0.00481, 0.00482,0.00483, 0.00484, 0.00485, 0.00486, 0.00487, 0.00488, 0.00489, 0.00490,0.00491, 0.00492, 0.00493, 0.00494, 0.00495, 0.00496, 0.00497, 0.00498,0.00499, 0.00500, 0.00501, 0.00502, 0.00503, 0.00504, 0.00505, 0.00506,0.00507, 0.00508, 0.00509, 0.00510, 0.00511, 0.00512, 0.00513, 0.00514,0.00515, 0.00516, 0.00517, 0.00518, 0.00519, 0.00520, 0.00521, 0.00522,0.00523, 0.00524, 0.00525, 0.00526, 0.00527, 0.00528, 0.00529, 0.00530,0.00531, 0.00532, 0.00533, 0.00534, 0.00535, 0.00536, 0.00537, 0.00538,0.00539, 0.00540, 0.00541, 0.00542, 0.00543, 0.00544, 0.00545, 0.00546,0.00547, 0.00548, 0.00549, 0.00550, 0.00551, 0.00552, 0.00553, 0.00554,0.00555, 0.00556, 0.00557, 0.00558, 0.00559, 0.00560, 0.00561, 0.00562,0.00563, 0.00564, 0.00565, 0.00566, 0.00567, 0.00568, 0.00569, 0.00570,0.00571, 0.00572, 0.00573, 0.00574, 0.00575, 0.00576, 0.00578, 0.00579,0.00580, 0.00581, 0.00582, 0.00583, 0.00584, 0.00585, 0.00586, 0.00587,0.00588, 0.00589, 0.00590, 0.00591, 0.00592, 0.00593, 0.00594, 0.00595,0.00596, 0.00597, 0.00598, 0.00599, 0.00600, 0.00601, 0.00602, 0.00603,0.00604, 0.00605, 0.00606, 0.00607, 0.00608, 0.00609, 0.00610, 0.00611,0.00612, 0.00613, 0.00614, 0.00615, 0.00616, 0.00617, 0.00618, 0.00619,0.00620, 0.00621, 0.00622, 0.00623, 0.00624, 0.00625, 0.00626, 0.00627,0.00628, 0.00629, 0.00630, 0.00631, 0.00632, 0.00633, 0.00634, 0.00635,0.00636, 0.00637, 0.00638, 0.00639, 0.00640, 0.00641, 0.00642, 0.00643,0.00644, 0.00645, 0.00646, 0.00647, 0.00648, 0.00649, 0.00650, 0.00651,0.00652, 0.00653, 0.00654, 0.00655, 0.00656, 0.00657, 0.00658, 0.00659,0.00660, 0.00661, 0.00662, 0.00663, 0.00664, 0.00665, 0.00666, 0.00667,0.00668, 0.00669, 0.00670, 0.00671, 0.00672, 0.00673, 0.00674, 0.00675,0.00676, 0.00678, 0.00679, 0.00680, 0.00681, 0.00682, 0.00683, 0.00684,0.00685, 0.00686, 0.00687, 0.00688, 0.00689, 0.00690, 0.00691, 0.00692,0.00693, 0.00694, 0.00695, 0.00696, 0.00697, 0.00698, 0.00699, 0.00700,0.00701, 0.00702, 0.00703, 0.00704, 0.00705, 0.00706, 0.00707, 0.00708,0.00709, 0.00710, 0.00711, 0.00712, 0.00713, 0.00714, 0.00715, 0.00716,0.00717, 0.00718, 0.00719, 0.00720, 0.00721, 0.00722, 0.00723, 0.00724,0.00725, 0.00726, 0.00727, 0.00728, 0.00729, 0.00730, 0.00731, 0.00732,0.00733, 0.00734, 0.00735, 0.00736, 0.00737, 0.00738, 0.00739, 0.00740,0.00741, 0.00742, 0.00743, 0.00744, 0.00745, 0.00746, 0.00747, 0.00748,0.00749, 0.00750, 0.00751, 0.00752, 0.00753, 0.00754, 0.00755, 0.00756,0.00757, 0.00758, 0.00759, 0.00760, 0.00761, 0.00762, 0.00763, 0.00764,0.00765, 0.00766, 0.00767, 0.00768, 0.00769, 0.00770, 0.00771, 0.00772,0.00773, 0.00774, 0.00775, 0.00776, 0.00777, 0.00778, 0.00779, 0.00780,0.00781, 0.00782, 0.00783, 0.00784, 0.00785, 0.00786, 0.00787, 0.00788,0.00789, 0.00790, 0.00791, 0.00792, 0.00793, 0.00794, 0.00795, 0.00796,0.00797, 0.00798, 0.00799, 0.00800, 0.00801, 0.00802, 0.00803, 0.00804,0.00805, 0.00806, 0.00807, 0.00808, 0.00809, 0.00810, 0.00811, 0.00812,0.00813, 0.00814, 0.00815, 0.00816, 0.00817, 0.00818, 0.00819, 0.00820,0.00821, 0.00822, 0.00823, 0.00824, 0.00825, 0.00826, 0.00827, 0.00828,0.00829, 0.00830, 0.00831, 0.00832, 0.00833, 0.00834, 0.00835, 0.00836,0.00837, 0.00838, 0.00839, 0.00840, 0.00841, 0.00842, 0.00843, 0.00844,0.00845, 0.00846, 0.00847, 0.00848, 0.00849, 0.00850, 0.00851, 0.00852,0.00853, 0.00854, 0.00855, 0.00856, 0.00857, 0.00858, 0.00859, 0.00860,0.00861, 0.00862, 0.00863, 0.00864, 0.00865, 0.00866, 0.00867, 0.00868,0.00869, 0.00870, 0.00871, 0.00872, 0.00873, 0.00874, 0.00875, 0.00876,0.00877, 0.00878, 0.00879, 0.00880, 0.00881, 0.00882, 0.00883, 0.00884,0.00885, 0.00886, 0.00887, 0.00888, 0.00889, 0.00890, 0.00891, 0.00892,0.00893, 0.00894, 0.00895, 0.00896, 0.00897, 0.00898, 0.00899, 0.00900,0.00901, 0.00902, 0.00903, 0.00904, 0.00905, 0.00906, 0.00907, 0.00908,0.00909, 0.00910, 0.00911, 0.00912, 0.00913, 0.00914, 0.00915, 0.00916,0.00917, 0.00918, 0.00919, 0.00920, 0.00921, 0.00922, 0.00923, 0.00924,0.00925, 0.00926, 0.00927, 0.00928, 0.00929, 0.00930, 0.00931, 0.00932,0.00933, 0.00934, 0.00935, 0.00936, 0.00937, 0.00938, 0.00939, 0.00940,0.00941, 0.00942, 0.00943, 0.00944, 0.00945, 0.00946, 0.00947, 0.00948,0.00949, 0.00950, 0.00951, 0.00952, 0.00953, 0.00954, 0.00955, 0.00956,0.00957, 0.00958, 0.00959, 0.00960, 0.00961, 0.00962, 0.00963, 0.00964,0.00965, 0.00966, 0.00967, 0.00968, 0.00969, 0.00970, 0.00971, 0.00972,0.00973, 0.00974, 0.00975, 0.00976, 0.00977, 0.00978, 0.00979, 0.00980,0.00981, 0.00982, 0.00983, 0.00984, 0.00985, 0.00986, 0.00987, 0.00988,0.00989, 0.00990, 0.00991, 0.00992, 0.00993, 0.00994, 0.00995, 0.00996,0.00997, 0.00998, 0.00999, 0.0100, 0.0101, 0.0103, 0.0104, 0.0105,0.0106, 0.0107, 0.0108, 0.0109, 0.0110, 0.0111, 0.0112, 0.0113, 0.0114,0.0115, 0.0116, 0.0117, 0.0118, 0.0119, 0.0120, 0.0121, 0.0122, 0.0123,0.0124, 0.0125, 0.0126, 0.0127, 0.0128, 0.0129, or 0.0130. In someembodiments, the ratio of CD34⁺ CD90⁺ CD45RA⁻ cells to monocytes in thesample is from about 0.0063 to about 0.0083, such as a ratio of CD34⁺CD90⁺ CD45RA⁻ cells to monocytes in the sample of about 0.00630,0.00631, 0.00632, 0.00633, 0.00634, 0.00635, 0.00636, 0.00637, 0.00638,0.00639, 0.00640, 0.00641, 0.00642, 0.00643, 0.00644, 0.00645, 0.00646,0.00647, 0.00648, 0.00649, 0.00650, 0.00651, 0.00652, 0.00653, 0.00654,0.00655, 0.00656, 0.00657, 0.00658, 0.00659, 0.00660, 0.00661, 0.00662,0.00663, 0.00664, 0.00665, 0.00666, 0.00667, 0.00668, 0.00669, 0.00670,0.00671, 0.00672, 0.00673, 0.00674, 0.00675, 0.00676, 0.00678, 0.00679,0.00680, 0.00681, 0.00682, 0.00683, 0.00684, 0.00685, 0.00686, 0.00687,0.00688, 0.00689, 0.00690, 0.00691, 0.00692, 0.00693, 0.00694, 0.00695,0.00696, 0.00697, 0.00698, 0.00699, 0.00700, 0.00701, 0.00702, 0.00703,0.00704, 0.00705, 0.00706, 0.00707, 0.00708, 0.00709, 0.00710, 0.00711,0.00712, 0.00713, 0.00714, 0.00715, 0.00716, 0.00717, 0.00718, 0.00719,0.00720, 0.00721, 0.00722, 0.00723, 0.00724, 0.00725, 0.00726, 0.00727,0.00728, 0.00729, 0.00730, 0.00731, 0.00732, 0.00733, 0.00734, 0.00735,0.00736, 0.00737, 0.00738, 0.00739, 0.00740, 0.00741, 0.00742, 0.00743,0.00744, 0.00745, 0.00746, 0.00747, 0.00748, 0.00749, 0.00750, 0.00751,0.00752, 0.00753, 0.00754, 0.00755, 0.00756, 0.00757, 0.00758, 0.00759,0.00760, 0.00761, 0.00762, 0.00763, 0.00764, 0.00765, 0.00766, 0.00767,0.00768, 0.00769, 0.00770, 0.00771, 0.00772, 0.00773, 0.00774, 0.00775,0.00776, 0.00777, 0.00778, 0.00779, 0.00780, 0.00781, 0.00782, 0.00783,0.00784, 0.00785, 0.00786, 0.00787, 0.00788, 0.00789, 0.00790, 0.00791,0.00792, 0.00793, 0.00794, 0.00795, 0.00796, 0.00797, 0.00798, 0.00799,0.00800, 0.00801, 0.00802, 0.00803, 0.00804, 0.00805, 0.00806, 0.00807,0.00808, 0.00809, 0.00810, 0.00811, 0.00812, 0.00813, 0.00814, 0.00815,0.00816, 0.00817, 0.00818, 0.00819, 0.00820, 0.00821, 0.00822, 0.00823,0.00824, 0.00825, 0.00826, 0.00827, 0.00828, 0.00829, 0.00830. In someembodiments, the ratio of CD34⁺ CD90⁺ CD45RA⁻ cells to monocytes in thesample is about 0.0073.

In an additional aspect, the invention features a method of mobilizing apopulation of hematopoietic stem cells from the bone marrow of amammalian donor (e.g., a human donor) into peripheral blood, the methodincluding administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to enrich the peripheral blood of thedonor with CD34⁺ CD90⁺ CD45RA⁻ cells relative to monocytes by a ratio offrom about 1.5:1 to about 8.5:1 as assessed by comparing a sample ofperipheral blood of the donor following administration of the CXCR2agonist and CXCR4 antagonist to a sample of peripheral blood of thedonor prior to administration of the CXCR2 agonist and CXCR4 antagonist.In some embodiments, the peripheral blood of the donor may be enrichedwith CD34⁺ CD90⁺ CD45RA⁻ cells relative to monocytes by a ratio of about1.50:1, 1.55:1, 1.60:1, 1.65:1, 1.70:1, 1.75:1, 1.80:1, 1.85:1, 1.90:1,1.95:1, 2.00:1, 2.05:1, 2.10:1, 2.15:1, 2.20:1, 2.25:1, 2.30:1, 2.35:1,2.40:1, 2.45:1, 2.50:1, 2.55:1, 2.60:1, 2.65:1, 2.70:1, 2.75:1, 2.80:1,2.85:1, 2.90:1, 2.95:1, 3.00:1, 3.05:1, 3.10:1, 3.15:1, 3.20:1, 3.25:1,3.30:1, 3.35:1, 3.40:1, 3.45:1, 3.50:1, 3.55:1, 3.60:1, 3.65:1, 3.70:1,3.75:1, 3.80:1, 3.85:1, 3.90:1, 3.95:1, 4.00:1, 4.05:1, 4.10:1, 4.15:1,4.20:1, 4.25:1, 4.30:1, 4.35:1, 4.40:1, 4.45:1, 4.50:1, 4.55:1, 4.60:1,4.65:1, 4.70:1, 4.75:1, 4.80:1, 4.85:1, 4.90:1, 4.95:1, 5.00:1, 5.05:1,5.10:1, 5.15:1, 5.20:1, 5.25:1, 5.30:1, 5.35:1, 5.40:1, 5.45:1, 5.50:1,5.55:1, 5.60:1, 5.65:1, 5.70:1, 5.75:1, 5.80:1, 5.85:1, 5.90:1, 5.95:1,6.00:1, 6.05:1, 6.10:1, 6.15:1, 6.20:1, 6.25:1, 6.30:1, 6.35:1, 6.40:1,6.45:1, 6.50:1, 6.55:1, 6.60:1, 6.65:1, 6.70:1, 6.75:1, 6.80:1, 6.85:1,6.90:1, 6.95:1, 7.00:1, 7.05:1, 7.10:1, 7.15:1, 7.20:1, 7.25:1, 7.30:1,7.35:1, 7.40:1, 7.45:1, 7.50:1, 7.55:1, 7.60:1, 7.65:1, 7.70:1, 7.75:1,7.80:1, 7.85:1, 7.90:1, 7.95:1, 8.00:1, 8.05:1, 8.10:1, 8.15:1, 8.20:1,8.25:1, 8.30:1, 8.35:1, 8.40:1, 8.45:1, or 8.50:1. In some embodiments,the peripheral blood of the donor is enriched with CD34⁺ CD90⁺ CD45RA⁻cells relative to monocytes by a ratio of about 1.9:1.

In another aspect, the invention features a method of mobilizing apopulation of hematopoietic stem cells from the bone marrow of amammalian donor (e.g., a human donor) into peripheral blood, the methodincluding administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to produce a population of cells havinga density of CD34⁺ CD90⁺ CD45RA⁻ cells of at least about 16,000cells/ml, such as a density of from about 20,000 cells/ml to about75,000 cells/ml, about 25,000 cells/ml to about 70,000 cells/ml, about30,000 cells/ml to about 65,000 cells/ml, about 35,000 cells/ml to about60,000 cells/ml, about 40,000 cells/ml to about 55,000 cells/ml, orabout 45,000 cells/ml to about 50,000 cells/ml (e.g., about 16,000cells/ml, 17,000 cells/ml, 18,000 cells/ml, 19,000 cells/ml, 20,000cells/ml, 21,000 cells/ml, 22,000 cells/ml, 23,000 cells/ml, 24,000cells/ml, 25,000 cells/ml, 26,000 cells/ml, 27,000 cells/ml, 28,000cells/ml, 29,000 cells/ml, 30,000 cells/ml, 31,000 cells/ml, 32,000cells/ml, 33,000 cells/ml, 34,000 cells/ml, 35,000 cells/ml, 36,000cells/ml, 37,000 cells/ml, 38,000 cells/ml, 39,000 cells/ml, 40,000cells/ml, 41,000 cells/nil, 42,000 cells/ml, 43,000 cells/ml, 44,000cells/ml, 45,000 cells/ml, 46,000 cells/ml, 47,000 cells/ml, 48,000cells/ml, 49,000 cells/ml, 50,000 cells/ml, 51,000 cells/ml, 52,000cells/ml, 53,000 cells/ml, 54,000 cells/ml, 55,000 cells/ml, 56,000cells/ml, 57,000 cells/ml, 58,000 cells/ml, 59,000 cells/ml, 60,000cells/ml, 61,000 cells/ml, 62,000 cells/ml, 63,000 cells/ml, 64,000cells/ml, 65,000 cells/ml, 66,000 cells/ml, 67,000 cells/ml, 68,000cells/ml, 69,000 cells/ml, 70,000 cells/ml, 71,000 cells/ml, 72,000cells/ml, 73,000 cells/ml, 74,000 cells/ml, 75,000 cells/ml, 76,000cells/ml, 77,000 cells/ml, or more), and having a density of monocytesof no more than about 6×10⁶ cells/ml, such as a density of monocytes offrom 3.4×10⁶ cells/ml to about 5.9×10⁶ cells/ml, about 3.5×10⁶ cells/mlto about 5.7×10⁶ cells/ml, or about 4×10⁶ cells/ml to about 5×10⁶cells/ml (e.g., 5.9×10⁶ cells/ml, 5.8×10⁶ cells/ml, 5.7×10⁶ cells/ml,5.6×10⁶ cells/ml, 5.5×10⁶ cells/ml, 5.4×10⁶ cells/ml, 5.3×10⁶ cells/ml,5.2×10⁶ cells/ml, 5.1×10⁶ cells/ml, 5×10⁶ cells/ml, 4.9×10⁶ cells/ml,4.8×10⁶ cells/ml, 4.7×10⁶ cells/ml, 4.6×10⁶ cells/ml, 4.5×10⁶ cells/ml,4.4×10⁶ cells/ml, 4.3×10⁶ cells/ml, 4.2×10⁶ cells/ml, 4.1×10⁶ cells/ml,4×10⁶ cells/ml, 3.9×10⁶ cells/ml, 3.8×10⁶ cells/ml, 3.7×10⁶ cells/ml,3.6×10⁶ cells/ml, 3.5×10⁶ cells/ml, 3.4×10⁶ cells/ml, or less). In someembodiments, the method includes administering to the donor a CXCR2agonist and a CXCR4 antagonist in amounts sufficient to produce apopulation of cells having a density of CD34⁺ CD90⁺ CD45RA⁻ cells offrom about 20,000 cells/ml to about 75,000 cells/ml, and having adensity of monocytes of from about 3.4×10⁶ cells/ml to about 6×10⁶cells/ml. In some embodiments, the method includes administering to thedonor a CXCR2 agonist and a CXCR4 antagonist in amounts sufficient toproduce a population of cells having a density of CD34⁺ CD90⁺ CD45RA⁻cells of from about 30,000 cells/ml to about 60,000 cells/ml, and havinga density of monocytes of from about 4×10⁶ cells/ml to about 5.5×10⁶cells/ml. In some embodiments, the method includes administering to thedonor a CXCR2 agonist and a CXCR4 antagonist in amounts sufficient toproduce a population of cells having a density of CD34⁺ CD90⁺ CD45RA⁻cells of from about 40,000 cells/ml to about 50,000 cells/ml, and havinga density of monocytes of from about 4×10⁶ cells/ml to about 5×10⁶cells/ml.

In another aspect, the invention features a method of mobilizing apopulation of hematopoietic stem cells from the bone marrow of amammalian donor (e.g., a human donor) into peripheral blood, the methodincluding administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to produce a population of cells havinga ratio of CD34⁺ CD90⁺ CD45RA⁻ cells to CD34⁺ cells of from about 0.393to about 0.745 in a sample of peripheral blood of the donor followingadministration of the CXCR2 agonist. In some embodiments, the ratio ofCD34⁺ CD90⁺ CD45RA⁻ cells to CD34⁺ cells in the sample may be about0.393, 0.394, 0.395, 0.396, 0.397, 0.398, 0.399, 0.401, 0.402, 0.403,0.404, 0.405, 0.406, 0.407, 0.408, 0.409, 0.410, 0.411, 0.412, 0.413,0.414, 0.415, 0.416, 0.417, 0.418, 0.419, 0.420, 0.421, 0.422, 0.423,0.424, 0.425, 0.426, 0.427, 0.428, 0.429, 0.430, 0.431, 0.432, 0.433,0.434, 0.435, 0.436, 0.437, 0.438, 0.439, 0.440, 0.441, 0.442, 0.443,0.444, 0.445, 0.446, 0.447, 0.448, 0.449, 0.450, 0.451, 0.452, 0.453,0.454, 0.455, 0.456, 0.457, 0.458, 0.459, 0.460, 0.461, 0.462, 0.463,0.464, 0.465, 0.466, 0.467, 0.468, 0.469, 0.470, 0.471, 0.472, 0.473,0.474, 0.475, 0.476, 0.478, 0.479, 0.480, 0.481, 0.482, 0.483, 0.484,0.485, 0.486, 0.487, 0.488, 0.489, 0.490, 0.491, 0.492, 0.493, 0.494,0.495, 0.496, 0.497, 0.498, 0.499, 0.500, 0.501, 0.502, 0.503, 0.504,0.505, 0.506, 0.507, 0.508, 0.509, 0.510, 0.511, 0.512, 0.513, 0.514,0.515, 0.516, 0.517, 0.518, 0.519, 0.520, 0.521, 0.522, 0.523, 0.524,0.525, 0.526, 0.527, 0.528, 0.529, 0.530, 0.531, 0.532, 0.533, 0.534,0.535, 0.536, 0.537, 0.538, 0.539, 0.540, 0.541, 0.542, 0.543, 0.544,0.545, 0.546, 0.547, 0.548, 0.549, 0.550, 0.551, 0.552, 0.553, 0.554,0.555, 0.556, 0.557, 0.558, 0.559, 0.560, 0.561, 0.562, 0.563, 0.564,0.565, 0.566, 0.567, 0.568, 0.569, 0.570, 0.571, 0.572, 0.573, 0.574,0.575, 0.576, 0.578, 0.579, 0.580, 0.581, 0.582, 0.583, 0.584, 0.585,0.586, 0.587, 0.588, 0.589, 0.590, 0.591, 0.592, 0.593, 0.594, 0.595,0.596, 0.597, 0.598, 0.599, 0.600, 0.601, 0.602, 0.603, 0.604, 0.605,0.606, 0.607, 0.608, 0.609, 0.610, 0.611, 0.612, 0.613, 0.614, 0.615,0.616, 0.617, 0.618, 0.619, 0.620, 0.621, 0.622, 0.623, 0.624, 0.625,0.626, 0.627, 0.628, 0.629, 0.630, 0.631, 0.632, 0.633, 0.634, 0.635,0.636, 0.637, 0.638, 0.639, 0.640, 0.641, 0.642, 0.643, 0.644, 0.645,0.646, 0.647, 0.648, 0.649, 0.650, 0.651, 0.652, 0.653, 0.654, 0.655,0.656, 0.657, 0.658, 0.659, 0.660, 0.661, 0.662, 0.663, 0.664, 0.665,0.666, 0.667, 0.668, 0.669, 0.670, 0.671, 0.672, 0.673, 0.674, 0.675,0.676, 0.678, 0.679, 0.680, 0.681, 0.682, 0.683, 0.684, 0.685, 0.686,0.687, 0.688, 0.689, 0.690, 0.691, 0.692, 0.693, 0.694, 0.695, 0.696,0.697, 0.698, 0.699, 0.700, 0.701, 0.702, 0.703, 0.704, 0.705, 0.706,0.707, 0.708, 0.709, 0.710, 0.711, 0.712, 0.713, 0.714, 0.715, 0.716,0.717, 0.718, 0.719, 0.720, 0.721, 0.722, 0.723, 0.724, 0.725, 0.726,0.727, 0.728, 0.729, 0.730, 0.731, 0.732, 0.733, 0.734, 0.735, 0.736,0.737, 0.738, 0.739, 0.740, 0.741, 0.742, 0.743, 0.744, or 0.745. Insome embodiments, the ratio of CD34⁺ CD90⁺ CD45RA⁻ cells to CD34⁺ cellsin the sample is from about 0.625 to about 0.725, such as a ratio ofCD34⁺ CD90⁺ CD45RA⁻ cells to CD34⁺ cells in the sample of about 0.625,0.626, 0.627, 0.628, 0.629, 0.630, 0.631, 0.632, 0.633, 0.634, 0.635,0.636, 0.637, 0.638, 0.639, 0.640, 0.641, 0.642, 0.643, 0.644, 0.645,0.646, 0.647, 0.648, 0.649, 0.650, 0.651, 0.652, 0.653, 0.654, 0.655,0.656, 0.657, 0.658, 0.659, 0.660, 0.661, 0.662, 0.663, 0.664, 0.665,0.666, 0.667, 0.668, 0.669, 0.670, 0.671, 0.672, 0.673, 0.674, 0.675,0.676, 0.678, 0.679, 0.680, 0.681, 0.682, 0.683, 0.684, 0.685, 0.686,0.687, 0.688, 0.689, 0.690, 0.691, 0.692, 0.693, 0.694, 0.695, 0.696,0.697, 0.698, 0.699, 0.700, 0.701, 0.702, 0.703, 0.704, 0.705, 0.706,0.707, 0.708, 0.709, 0.710, 0.711, 0.712, 0.713, 0.714, 0.715, 0.716,0.717, 0.718, 0.719, 0.720, 0.721, 0.722, 0.723, 0.724, or 0.725. Insome embodiments, the ratio of CD34⁺ CD90⁺ CD45RA⁻ cells to CD34⁺ cellsin the sample is about 0.676.

In an additional aspect, the invention features a method of mobilizing apopulation of hematopoietic stem cells from the bone marrow of amammalian donor (e.g., a human donor) into peripheral blood, the methodincluding administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to enrich the peripheral blood of thedonor with CD34⁺ CD90⁺ CD45RA⁻ cells relative to CD34⁺ cells by a ratioof from about 1.1:1 to about 4.8:1 as assessed by comparing a sample ofperipheral blood of the donor following administration of the CXCR2agonist and CXCR4 antagonist to a sample of peripheral blood of thedonor prior to administration of the CXCR2 agonist and CXCR4 antagonist.In some embodiments, the peripheral blood of the donor may be enrichedwith CD34⁺ CD90⁺ CD45RA⁻ cells relative to monocytes by a ratio of about1.10:1, 1.15:1, 1.20:1, 1.25:1, 1.30:1, 1.35:1, 1.40:1, 1.45:1, 1.50:1,1.55:1, 1.60:1, 1.65:1, 1.70:1, 1.75:1, 1.80:1, 1.85:1, 1.90:1, 1.95:1,2.00:1, 2.05:1, 2.10:1, 2.15:1, 2.20:1, 2.25:1, 2.30:1, 2.35:1, 2.40:1,2.45:1, 2.50:1, 2.55:1, 2.60:1, 2.65:1, 2.70:1, 2.75:1, 2.80:1, 2.85:1,2.90:1, 2.95:1, 3.00:1, 3.05:1, 3.10:1, 3.15:1, 3.20:1, 3.25:1, 3.30:1,3.35:1, 3.40:1, 3.45:1, 3.50:1, 3.55:1, 3.60:1, 3.65:1, 3.70:1, 3.75:1,3.80:1, 3.85:1, 3.90:1, 3.95:1, 4.00:1, 4.05:1, 4.10:1, 4.15:1, 4.20:1,4.25:1, 4.30:1, 4.35:1, 4.40:1, 4.45:1, 4.50:1, 4.55:1, 4.60:1, 4.65:1,4.70:1, 4.75:1, or 4.80:1. In some embodiments, the peripheral blood ofthe donor is enriched with CD34⁺ CD90⁺ CD45RA⁻ cells relative to CD34⁺cells by a ratio of about 1.2:1.

In another aspect, the invention features a method of mobilizing apopulation of hematopoietic stem cells from the bone marrow of amammalian donor (e.g., a human donor) into peripheral blood, the methodincluding administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to produce a population of cells havinga frequency of CD34⁺ CD90⁺ CD45RA⁻ cells of from about 0.020% to about0.110% in a sample of peripheral blood of the donor followingadministration of the CXCR2 agonist and CXCR4 antagonist. In someembodiments, the population of cells may have a frequency of CD34⁺ CD90⁺CD45RA⁻ cells of about 0.020%, 0.021%, 0.022%, 0.023%, 0.024%, 0.025%,0.026%, 0.027%, 0.028%, 0.029%, 0.030%, 0.031%, 0.032%, 0.033%, 0.034%,0.035%, 0.036%, 0.037%, 0.038%, 0.039%, 0.040%, 0.041%, 0.042%, 0.043%,0.044%, 0.045%, 0.046%, 0.047%, 0.048%, 0.049%, 0.050%, 0.051%, 0.052%,0.053%, 0.054%, 0.055%, 0.056%, 0.057%, 0.058%, 0.059%, 0.060%, 0.061%,0.062%, 0.063%, 0.064%, 0.065%, 0.066%, 0.067%, 0.068%, 0.069%, 0.070%,0.071%, 0.072%, 0.073%, 0.074%, 0.075%, 0.076%, 0.077%, 0.078%, 0.079%,0.080%, 0.081%, 0.082%, 0.083%, 0.084%, 0.085%, 0.086%, 0.087%, 0.088%,0.089%, 0.090%, 0.091%, 0.092%, 0.093%, 0.094%, 0.095%, 0.096%, 0.097%,0.098%, 0.099%, 0.100%, 0.101%, 0.102%, 0.103%, 0.104%, 0.105%, 0.106%,0.107%, 0.108%, 0.109%, or 0.110%. In some embodiments, the populationof cells has a frequency of CD34⁺ CD90⁺ CD45RA⁻ cells of from about0.046% to about 0.086%, such as a frequency of hematopoietic stem cellsof about 0.046%, 0.047%, 0.048%, 0.049%, 0.050%, 0.051%, 0.052%, 0.053%,0.054%, 0.055%, 0.056%, 0.057%, 0.058%, 0.059%, 0.060%, 0.061%, 0.062%,0.063%, 0.064%, 0.065%, 0.066%, 0.067%, 0.068%, 0.069%, 0.070%, 0.071%,0.072%, 0.073%, 0.074%, 0.075%, 0.076%, 0.077%, 0.078%, 0.079%, 0.080%,0.081%, 0.082%, 0.083%, 0.084%, 0.085%, or 0.086%. In some embodiments,the population of cells has a frequency of CD34⁺ CD90⁺ CD45RA⁻ cells ofabout 0.066%.

In an additional aspect, the invention features a method of mobilizing apopulation of hematopoietic stem cells from the bone marrow of amammalian donor (e.g., a human donor) into peripheral blood, the methodincluding administering to the donor a CXCR2 agonist and a CXCR4antagonist in amounts sufficient to induce an increase in the frequencyof CD34⁺ CD90⁺ CD45RA⁻ cells in the peripheral blood of the donor by atleast 3-fold as assessed by comparing a sample of peripheral blood ofthe donor following administration of the CXCR2 agonist and CXCR4antagonist to a sample of peripheral blood of the donor prior toadministration of the CXCR2 agonist and CXCR4 antagonist (e.g., by fromabout 5.1-fold to about 25.7-fold, such as by about 5.1-fold, 5.2-fold,5.3-fold, 5.4-fold, 5.5-fold, 5.6-fold, 5.7-fold, 5.8-fold, 5.9-fold,6.0-fold, 6.1-fold, 6.2-fold, 6.3-fold, 6.4-fold, 6.5-fold, 6.6-fold,6.7-fold, 6.8-fold, 6.9-fold, 7.0-fold, 7.1-fold, 7.2-fold, 7.3-fold,7.4-fold, 7.5-fold, 7.6-fold, 7.7-fold, 7.8-fold, 7.9-fold, 8.0-fold,8.1-fold, 8.2-fold, 8.3-fold, 8.4-fold, 8.5-fold, 8.6-fold, 8.7-fold,8.8-fold, 8.9-fold, 9.0-fold, 9.1-fold, 9.2-fold, 9.3-fold, 9.4-fold,9.5-fold, 9.6-fold, 9.7-fold, 9.8-fold, 9.9-fold, 10.0-fold, 10.1-fold,10.2-fold, 10.3-fold, 10.4-fold, 10.5-fold, 10.6-fold, 10.7-fold,10.8-fold, 10.9-fold, 11.0-fold, 11.1-fold, 11.2-fold, 11.3-fold,11.4-fold, 11.5-fold, 11.6-fold, 11.7-fold, 11.8-fold, 11.9-fold,12.0-fold, 12.1-fold, 12.2-fold, 12.3-fold, 12.4-fold, 12.5-fold,12.6-fold, 12.7-fold, 12.8-fold, 12.9-fold, 13.0-fold, 13.1-fold,13.2-fold, 13.3-fold, 13.4-fold, 13.5-fold, 13.6-fold, 13.7-fold,13.8-fold, 13.9-fold, 14.0-fold, 14.1-fold, 14.2-fold, 14.3-fold,14.4-fold, 14.5-fold, 14.6-fold, 14.7-fold, 14.8-fold, 14.9-fold,15.0-fold, 15.1-fold, 15.2-fold, 15.3-fold, 15.4-fold, 15.5-fold,15.6-fold, 15.7-fold, 15.8-fold, 15.9-fold, 16.0-fold, 16.1-fold,16.2-fold, 16.3-fold, 16.4-fold, 16.5-fold, 16.6-fold, 16.7-fold,16.8-fold, 16.9-fold, 17.0-fold, 17.1-fold, 17.2-fold, 17.3-fold,17.4-fold, 17.5-fold, 17.6-fold, 17.7-fold, 17.8-fold, 17.9-fold,18.0-fold, 18.1-fold, 18.2-fold, 18.3-fold, 18.4-fold, 18.5-fold,18.6-fold, 18.7-fold, 18.8-fold, 18.9-fold, 19.0-fold, 19.1-fold,19.2-fold, 19.3-fold, 19.4-fold, 19.5-fold, 19.6-fold, 19.7-fold,19.8-fold, 19.9-fold, 20.0-fold, 20.1-fold, 20.2-fold, 20.3-fold,20.4-fold, 20.5-fold, 20.6-fold, 20.7-fold, 20.8-fold, 20.9-fold,21.0-fold, 21.1-fold, 21.2-fold, 21.3-fold, 21.4-fold, 21.5-fold,21.6-fold, 21.7-fold, 21.8-fold, 21.9-fold, 22.0-fold, 22.1-fold,22.2-fold, 22.3-fold, 22.4-fold, 22.5-fold, 22.6-fold, 22.7-fold,22.8-fold, 22.9-fold, 23.0-fold, 23.1-fold, 23.2-fold, 23.3-fold,23.4-fold, 23.5-fold, 23.6-fold, 23.7-fold, 23.8-fold, 23.9-fold,24.0-fold, 24.1-fold, 24.2-fold, 24.3-fold, 24.4-fold, 24.5-fold,24.6-fold, 24.7-fold, 24.8-fold, 24.9-fold, 25.0-fold, 25.1-fold,25.2-fold, 25.3-fold, 25.4-fold, 25.5-fold, 25.6-fold, or 25.7-fold. Insome embodiments, the frequency of CD34⁺ CD90⁺ CD45RA⁻ cells in theperipheral blood of the donor is increased by from about 5.1-fold toabout 7.1-fold following administration of the CXCR2 agonist and CXCR4antagonist, such as by about 5.1-fold, 5.2-fold, 5.3-fold, 5.4-fold,5.5-fold, 5.6-fold, 5.7-fold, 5.8-fold, 5.9-fold, 6.0-fold, 6.1-fold,6.2-fold, 6.3-fold, 6.4-fold, 6.5-fold, 6.6-fold, 6.7-fold, 6.8-fold,6.9-fold, 7.0-fold, or 7.1-fold. In some embodiments, the frequency ofCD34⁺ CD90⁺ CD45RA⁻ cells in the peripheral blood of the donor isincreased by about 5.8-fold.

In another aspect, the invention features a method of mobilizing apopulation of hematopoietic stem cells, from the bone marrow of amammalian donor into peripheral blood, the method comprisingadministering to the donor mobilizing amounts of a CXCR2 agonist and aCXCR4 antagonist; acquiring an input value for each of one or moreparameters in TABLE 2 characterizing a sample of peripheral blood of thedonor, and releasing the sample for ex vivo expansion of thehematopoietic stem cells or for use in the treatment of one or more stemcell disorders in a mammalian patient if the input value for each of theone or more parameters meets the corresponding reference criterion foreach of the one or more parameters. In some embodiments, the one or morereference parameters are a set of parameters listed in any one of TABLES3-6 herein.

In some embodiments of any of the above aspects of the invention, thesample is isolated from the donor at from about 3 hours to about 5 hoursfollowing administration of the CXCR2 agonist and CXCR4 antagonist(e.g., at about 3 hours, 3.1 hours, 3.2 hours, 3.3 hours, 3.4 hours, 3.5hours, 3.6 hours, 3.7 hours, 3.8 hours, 3.9 hours, 4.0 hours, 4.1 hours,4.2 hours, 4.3 hours, 4.4 hours, 4.5 hours, 4.6 hours, 4.7 hours, 4.8hours, 4.9 hours, or 5.0 hours following administration of the CXCR2agonist and CXCR4 antagonist). In some embodiments, the sample isisolated from the donor at about 4 hours following administration of theCXCR2 agonist and CXCR4 antagonist.

In some embodiments of any of the above aspects of the invention, theCXCR2 agonist is Gro-β T or a variant thereof. In some embodiments, theCXCR2 agonist may be a peptide having at least about 85% (e.g., at leastabout 85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity tothe amino acid sequence of SEQ ID NO: 2. In some embodiments, the CXCR2agonist is a peptide having from about 85% to 100% sequence identity tothe amino acid sequence of SEQ ID NO: 2, such as a peptide having fromabout 86% to about 100%, from about 87% to about 99%, about 88% to about98%, about 89%, to about 97%, about 90% to about 96%, or about 91% toabout 95% sequence identity to the amino acid sequence of SEQ ID NO: 2.In some embodiments, the CXCR2 agonist is a peptide having an amino acidsequence that differs from that of SEQ ID NO: 2 only by way of one ormore conservative amino acid substitutions (e.g., only by way of from 1to 10 conservative amino acid substitutions, from 1 to 5 conservativeamino acid substitutions, or from 1 to 3 conservative amino acidsubstitutions, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 conservativeamino acid substitutions). In some embodiments, the CXCR2 agonist isGro-β T. In some embodiments, the Gro-β T is not covalently modified. Insome embodiments, the Gro-β T is not covalently modified with apolyalkylene glycol moiety, such as a polyethylene glycol moiety.

In some embodiments of any of the above aspects of the invention, theCXCR2 agonist is Gro-β or a variant thereof. In some embodiments, theCXCR2 agonist may be a peptide having at least about 85% (e.g., about85%, 90%, 95%, 96%, 97%, 98%, 99%, or more) sequence identity to theamino acid sequence of SEQ ID NO: 1. In some embodiments, the CXCR2agonist is a peptide having from about 85% to 100% sequence identity tothe amino acid sequence of SEQ ID NO: 1, such as a peptide having fromabout 86% to about 100%, from about 87% to about 99%, about 88% to about98%, about 89%, to about 97%, about 90% to about 96%, or about 91% toabout 95% sequence identity to the amino acid sequence of SEQ ID NO: 1.In some embodiments, the CXCR2 agonist is a peptide having an amino acidsequence that differs from that of SEQ ID NO: 1 only by way of one ormore conservative amino acid substitutions (e.g., only by way of from 1to 10 conservative amino acid substitutions, from 1 to 5 conservativeamino acid substitutions, or from 1 to 3 conservative amino acidsubstitutions, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 conservativeamino acid substitutions). In some embodiments, the CXCR2 agonist isGro-β. In some embodiments, the Gro-β T is not covalently modified. Insome embodiments, the Gro-β is not covalently modified with apolyalkylene glycol moiety, such as a polyethylene glycol moiety.

In some embodiments, the CXCR2 agonist (e.g., Gro-β or Gro-β T, such asunmodified Gro-β or Gro-β T) is administered to the donor at a dose offrom about 50 μg/kg to about 1 mg/kg, such as a dose of about 50 μg/kg,55 μg/kg, 60 μg/kg, 65 μg/kg, 70 μg/kg, 75 μg/kg, 80 μg/kg, 85 μg/kg, 90μg/kg, 95 μg/kg, 100 μg/kg, 105 μg/kg, 110 μg/kg, 115 μg/kg, 120 μg/kg,125 μg/kg, 130 μg/kg, 135 μg/kg, 140 μg/kg, 145 μg/kg, 150 μg/kg, 155μg/kg, 160 μg/kg, 165 μg/kg, 170 μg/kg, 175 μg/kg, 180 μg/kg, 185 μg/kg,190 μg/kg, 195 μg/kg, 200 μg/kg, 205 μg/kg, 210 μg/kg, 215 μg/kg, 220μg/kg, 225 μg/kg, 230 μg/kg, 235 μg/kg, 240 μg/kg, 245 μg/kg, 250 μg/kg,255 μg/kg, 260 μg/kg, 265 μg/kg, 270 μg/kg, 275 μg/kg, 280 μg/kg, 285μg/kg, 290 μg/kg, 295 μg/kg, 300 μg/kg, 305 μg/kg, 310 μg/kg, 315 μg/kg,320 μg/kg, 325 μg/kg, 330 μg/kg, 335 μg/kg, 340 μg/kg, 345 μg/kg, 350μg/kg, 355 μg/kg, 360 μg/kg, 365 μg/kg, 370 μg/kg, 375 μg/kg, 380 μg/kg,400 μg/kg, 405 μg/kg, 410 μg/kg, 415 μg/kg, 425 μg/kg, 430 μg/kg, 435μg/kg, 440 μg/kg, 445 μg/kg, 450 μg/kg, 210 μg/kg, 300 μg/kg, 400 μg/kg,405 μg/kg, 410 μg/kg, 415 μg/kg, 420 μg/kg, 425 μg/kg, 430 μg/kg, 435μg/kg, 440 μg/kg, 445 μg/kg, 450 μg/kg, 455 μg/kg, 460 μg/kg, 465 μg/kg,470 μg/kg, 475 μg/kg, 480 μg/kg, 485 μg/kg, 490 μg/kg, 495 μg/kg, 500μg/kg, 505 μg/kg, 510 μg/kg, 505 μg/kg, 515 μg/kg, 520 μg/kg, 525 μg/kg,530 μg/kg, 545 μg/kg, 550 μg/kg, 555 μg/kg, 560 μg/kg, 565 μg/kg, 570μg/kg, 575 μg/kg, 580 μg/kg, 585 μg/kg, 590 μg/kg, 595 μg/kg, 600 μg/kg,605 μg/kg, 610 μg/kg, 615 μg/kg, 620 μg/kg, 625 μg/kg, 630 μg/kg, 635μg/kg, 640 μg/kg, 645 μg/kg, 650 μg/kg, 655 μg/kg, 660 μg/kg, 665 μg/kg,670 μg/kg, 675 μg/kg, 680 μg/kg, 685 μg/kg, 690 μg/kg, 695 μg/kg, 700μg/kg, 705 μg/kg, 710 μg/kg, 715 μg/kg, 720 μg/kg, 725 μg/kg, 730 μg/kg,735 μg/kg, 740 μg/kg, 745 μg/kg, 750 μg/kg, 755 μg/kg, 760 μg/kg, 765μg/kg, 770 μg/kg, 775 μg/kg, 780 μg/kg, 785 μg/kg, 790 μg/kg, 795 μg/kg,800 μg/kg, 805 μg/kg, 810 μg/kg, 815 μg/kg, 820 μg/kg, 825 μg/kg, 830μg/kg, 835 μg/kg, 840 μg/kg, 845 μg/kg, 850 μg/kg, 855 μg/kg, 860 μg/kg,865 μg/kg, 870 μg/Kg, 875 μg/kg, 880 μg/kg, 885 μg/kg, 890 μg/kg, 895μg/kg, 900 μg/kg, 905 μg/kg, 910 μg/kg, 915 μg/kg, 920 μg/kg, 925 μg/kg,930 μg/kg, 935 μg/kg, 940 μg/kg, 945 μg/kg, 950 μg/kg, 955 μg/kg, 960μg/kg, 965 μg/kg, 970 μg/kg, 975 μg/kg, 980 μg/kg, 985 μg/kg, 990 μg/kg,995 μg/kg, or 1,000 μg/kg. In some embodiments, the CXCR2 agonist (e.g.,Gro-β or Gro-β T, such as unmodified Gro-β or Gro-β T) is administeredto the donor at a dose of from about 50 μg/kg to about 300 μg/kg, suchas a dose of from about 100 μg/kg to about 250 μg/kg, or from about 125μg/kg to about 225 μg/kg. In some embodiments, the CXCR2 agonist (e.g.,Gro-β or Gro-β T, such as unmodified Gro-β or Gro-β T) is administeredto the donor at a dose of about 150 μg/kg.

In another aspect, the invention features a method of mobilizing apopulation of hematopoietic stem cells from the bone marrow of amammalian donor (e.g., a human donor) into peripheral blood, the methodincluding administering to the donor a CXCR2 agonist selected from thegroup consisting of Gro-β, Gro-β T, and variants thereof at a dose offrom about 50 μg/kg to about 1 mg/kg (e.g., a dose of 50 μg/kg, 55μg/kg, 60 μg/kg, 65 μg/kg, 70 μg/kg, 75 μg/kg, 80 μg/kg, 85 μg/kg, 90μg/kg, 95 μg/kg, 100 μg/kg, 105 μg/kg, 110 μg/kg, 115 μg/kg, 120 μg/kg,125 μg/kg, 130 μg/kg, 135 μg/kg, 140 μg/kg, 145 μg/kg, 150 μg/kg, 155μg/kg, 160 μg/kg, 165 μg/kg, 170 μg/kg, 175 μg/kg, 180 μg/kg, 185 μg/kg,190 μg/kg, 195 μg/kg, 200 μg/kg, 205 μg/kg, 210 μg/kg, 215 μg/kg, 220μg/kg, 225 μg/kg, 230 μg/kg, 235 μg/kg, 240 μg/kg, 245 μg/kg, 250 μg/kg,255 μg/kg, 260 μg/kg, 265 μg/kg, 270 μg/kg, 275 μg/kg, 280 μg/kg, 285μg/kg, 290 μg/kg, 295 μg/kg, 300 μg/kg, 305 μg/kg, 310 μg/kg, 315 μg/kg,320 μg/kg, 325 μg/kg, 330 μg/kg, 335 μg/kg, 340 μg/kg, 345 μg/kg, 350μg/kg, 355 μg/kg, 360 μg/kg, 365 μg/kg, 370 μg/kg, 375 μg/kg, 380 μg/kg,400 μg/kg, 405 μg/kg, 410 μg/kg, 415 μg/kg, 425 μg/kg, 430 μg/kg, 435μg/kg, 440 μg/kg, 445 μg/kg, 450 μg/kg, 210 μg/kg, 300 μg/kg, 400 μg/kg,405 μg/kg, 410 μg/kg, 415 μg/kg, 420 μg/kg, 425 μg/kg, 430 μg/kg, 435μg/kg, 440 μg/kg, 445 μg/kg, 450 μg/kg, 455 μg/kg, 460 μg/kg, 465 μg/kg,470 μg/kg, 475 μg/kg, 480 μg/kg, 485 μg/kg, 490 μg/kg, 495 μg/kg, 500μg/kg, 505 μg/kg, 510 μg/kg, 505 μg/kg, 515 μg/kg, 520 μg/kg, 525 μg/kg,530 μg/kg, 545 μg/kg, 550 μg/kg, 555 μg/kg, 560 μg/kg, 565 μg/kg, 570μg/kg, 575 μg/kg, 580 μg/kg, 585 μg/kg, 590 μg/kg, 595 μg/kg, 600 μg/kg,605 μg/kg, 610 μg/kg, 615 μg/kg, 620 μg/kg, 625 μg/kg, 630 μg/kg, 635μg/kg, 640 μg/kg, 645 μg/kg, 650 μg/kg, 655 μg/kg, 660 μg/kg, 665 μg/kg,670 μg/kg, 675 μg/kg, 680 μg/kg, 685 μg/kg, 690 μg/kg, 695 μg/kg, 700μg/kg, 705 μg/kg, 710 μg/kg, 715 μg/kg, 720 μg/kg, 725 μg/kg, 730 μg/kg,735 μg/kg, 740 μg/kg, 745 μg/kg, 750 μg/kg, 755 μg/kg, 760 μg/kg, 765μg/kg, 770 μg/kg, 775 μg/kg, 780 μg/kg, 785 μg/kg, 790 μg/kg, 795 μg/kg,800 μg/kg, 805 μg/kg, 810 μg/kg, 815 μg/kg, 820 μg/kg, 825 μg/kg, 830μg/kg, 835 μg/kg, 840 μg/kg, 845 μg/kg, 850 μg/kg, 855 μg/kg, 860 μg/kg,865 μg/kg, 870 μg/Kg, 875 μg/kg, 880 μg/kg, 885 μg/kg, 890 μg/kg, 895μg/kg, 900 μg/kg, 905 μg/kg, 910 μg/kg, 915 μg/kg, 920 μg/kg, 925 μg/kg,930 μg/kg, 935 μg/kg, 940 μg/kg, 945 μg/kg, 950 μg/kg, 955 μg/kg, 960μg/kg, 965 μg/kg, 970 μg/kg, 975 μg/kg, 980 μg/kg, 985 μg/kg, 990 μs/kg,995 μg/kg, or 1,000 μg/kg). In some embodiments, the method furtherincludes administering a CXCR4 antagonist to the donor.

In some embodiments of any of the above aspects of the invention, theCXCR2 agonist (e.g., Gro-β or Gro-β T, such as unmodified Gro-β or Gro-βT) is administered to the donor at a dose of from about 50 μg/kg toabout 300 μg/kg, such as a dose of about 50 μg/kg, 55 μg/kg, 60 μg/kg,65 μg/kg, 70 μg/kg, 75 μg/kg, 80 μg/kg, 85 μg/kg, 90 μg/kg, 95 μg/kg,100 μg/kg, 105 μg/kg, 110 μg/kg, 115 μg/kg, 120 μg/kg, 125 μg/kg, 130μg/kg, 135 μg/kg, 140 μg/kg, 145 μg/kg, 150 μg/kg, 155 μg/kg, 160 μg/kg,165 μg/kg, 170 μg/kg, 175 μg/kg, 180 μg/kg, 185 μg/kg, 190 μg/kg, 195μg/kg, 200 μg/kg, 205 μg/kg, 210 μg/kg, 215 μg/kg, 220 μg/kg, 225 μg/kg,230 μg/kg, 235 μg/kg, 240 μg/kg, 245 μg/kg, 250 μg/kg, 255 μg/kg, 260μg/kg, 265 μg/kg, 270 μg/kg, 275 μg/kg, 280 μg/kg, 285 μg/kg, 290 μg/kg,295 μg/kg, or 300 μg/kg.

In some embodiments of any of the above aspects of the invention, theCXCR2 agonist (e.g., Gro-β or Gro-β T, such as unmodified Gro-β or Gro-βT) is administered to the donor at a dose of from about 100 μg/kg toabout 250 μg/kg, such as a dose of about 100 pig/kg, 105 μg/kg, 110μg/kg, 115 μg/kg, 120 μg/kg, 125 μg/kg, 130 μg/kg, 135 μg/kg, 140 μg/kg,145 μg/kg, 150 μg/kg, 155 μg/kg, 160 μg/kg, 165 μg/kg, 170 μg/kg, 175μg/kg, 180 μg/kg, 185 μg/kg, 190 μg/kg, 195 μg/kg, 200 μg/kg, 205 μg/kg,210 μg/kg, 215 μg/kg, 220 μg/kg, 225 μg/kg, 230 μg/kg, 235 μg/kg, 240μg/kg, 245 μg/kg, or 250 μg/kg.

In some embodiments of any of the above aspects of the invention, theCXCR2 agonist (e.g., Gro-β or Gro-β T, such as unmodified Gro-β or Gro-βT) is administered to the donor at a dose of about 150 μg/kg. Forexample, in some embodiments, the CXCR2 agonist (e.g., Gro-β or Gro-β T,such as unmodified Gro-β or Gro-β T) is administered to the donor at adose of from about 50 μg/kg per day to about 1 mg/kg per day, such as adose of about 50 μg/kg per day, 55 μg/kg per day, 60 μg/kg per day, 65μg/kg per day, 70 μg/kg per day, 75 μg/kg per day, 80 μg/kg per day, 85μg/kg per day, 90 μg/kg per day, 95 μg/kg per day, 100 pig/kg per day,105 μg/kg per day, 110 μg/kg per day, 115 μg/kg per day, 120 μg/kg perday, 125 μg/kg per day, 130 μg/kg per day, 135 μg/kg per day, 140 μg/kgper day, 145 μg/kg per day, 150 μg/kg per day, 155 μg/kg per day, 160μg/kg per day, 165 μg/kg per day, 170 μg/kg per day, 175 μg/kg per day,180 μg/kg per day, 185 μg/kg per day, 190 μg/kg per day, 195 pig/kg perday, 200 μg/kg per day, 205 μg/kg per day, 210 μg/kg per day, 215 μg/kgper day, 220 μg/kg per day, 225 μg/kg per day, 230 μg/kg per day, 235μg/kg per day, 240 μg/kg per day, 245 μg/kg per day, 250 μg/kg per day,255 μg/kg per day, 260 μg/kg per day, 265 μg/kg per day, 270 μg/kg perday, 275 μg/kg per day, 280 μg/kg per day, 285 μg/kg per day, 290 pig/kgper day, 295 μg/kg per day, 300 μg/kg per day, 305 μg/kg per day, 310μg/kg per day, 315 μg/kg per day, 320 μg/kg per day, 325 μg/kg per day,330 μg/kg per day, 335 μg/kg per day, 340 μg/kg per day, 345 μg/kg perday, 350 μg/kg per day, 355 μg/kg per day, 360 μg/kg per day, 365 μg/kgper day, 370 μg/kg per day, 375 μg/kg per day, 380 μg/kg per day, 400μg/kg per day, 405 μg/kg per day, 410 μg/kg per day, 415 μg/kg per day,425 μg/kg per day, 430 μg/kg per day, 435 μg/kg per day, 440 μg/kg perday, 445 μg/kg per day, 450 μg/kg per day, 210 μg/kg per day, 300 μg/kgper day, 400 μg/kg per day, 405 μg/kg per day, 410 μg/kg per day, 415μg/kg per day, 420 μg/kg per day, 425 μg/kg per day, 430 μg/kg per day,435 μg/kg per day, 440 μg/kg per day, 445 μg/kg per day, 450 μg/kg perday, 455 μg/kg per day, 460 μg/kg per day, 465 μg/kg per day, 470 μg/kgper day, 475 μg/kg per day, 480 μg/kg per day, 485 μg/kg per day, 490μg/kg per day, 495 μg/kg per day, 500 μg/kg per day, 505 μg/kg per day,510 μg/kg per day, 505 μg/kg per day, 515 μg/kg per day, 520 μg/kg perday, 525 Kg/kg per day, 530 μg/kg per day, 545 μg/kg per day, 550 μg/kgper day, 555 μg/kg per day, 560 μg/kg per day, 565 μg/kg per day, 570μg/kg per day, 575 μg/kg per day, 580 μg/kg per day, 585 μg/kg per day,590 μg/kg per day, 595 μg/kg per day, 600 μg/kg per day, 605 μg/kg perday, 610 μg/kg per day, 615 μg/kg per day, 620 μg/kg per day, 625 μg/kgper day, 630 μg/kg per day, 635 μg/kg per day, 640 μg/kg per day, 645μg/kg per day, 650 μg/kg per day, 655 μg/kg per day, 660 μg/kg per day,665 μg/kg per day, 670 μg/kg per day, 675 μg/kg per day, 680 μg/kg perday, 685 μg/kg per day, 690 μg/kg per day, 695 μg/kg per day, 700 μg/kgper day, 705 μg/kg per day, 710 μg/kg per day, 715 μg/kg per day, 720μg/kg per day, 725 μg/kg per day, 730 μg/kg per day, 735 μg/kg per day,740 μg/kg per day, 745 μg/kg per day, 750 μg/kg per day, 755 μg/kg perday, 760 μg/kg per day, 765 μg/kg per day, 770 μg/kg per day, 775 μg/kgper day, 780 μg/kg per day, 785 μg/kg per day, 790 μg/kg per day, 795μg/kg per day, 800 μg/kg per day, 805 μg/kg per day, 810 μg/kg per day,815 μg/kg per day, 820 pig/kg per day, 825 μg/kg per day, 830 μg/kg perday, 835 μg/kg per day, 840 μg/kg per day, 845 μg/kg per day, 850 μg/kgper day, 855 μg/kg per day, 860 μg/kg per day, 865 μg/kg per day, 870μg/kg per day, 875 μg/kg per day, 880 μg/kg per day, 885 μg/kg per day,890 μg/kg per day, 895 μg/kg per day, 900 μg/kg per day, 905 μg/kg perday, 910 μg/kg per day, 915 pig/kg per day, 920 μg/kg per day, 925 μg/kgper day, 930 μg/kg per day, 935 μg/kg per day, 940 μg/kg per day, 945μg/kg per day, 950 μg/kg per day, 955 μg/kg per day, 960 μg/kg per day,965 μg/kg per day, 970 μg/kg per day, 975 μg/kg per day, 980 μg/kg perday, 985 μg/kg per day, 990 μg/kg per day, 995 μg/kg per day, or 1,000μg/kg per day. In some embodiments, the CXCR2 agonist (e.g., Gro-β orGro-β T, such as unmodified Gro-β or Gro-β T) is administered to thedonor at a dose of from about 50 μg/kg per day to about 300 μg/kg perday, such as a dose of from about 100 μg/kg per day to about 250 μg/kgper day, or from about 125 μg/kg per day to about 225 μg/kg per day, orfrom about 125 μg/kg per day to about 175 μg/kg per day. In someembodiments, the CXCR2 agonist (e.g., Gro-β or Gro-β T, such asunmodified Gro-β or Gro-β T) is administered to the donor at a dose ofabout 150 μg/kg per day. In some embodiments, the CXCR2 agonist may beadministered as a single dose. In other embodiments, the CXCR2 agonistmay be administered as two or more doses.

In some embodiments, a human equivalent dose (HED) may be derived fromanimal dosage data using a conversion factor. For example, Nair andJacob, J. Basic Clin. Pharma. (2016) 7:27-31 disclose methodsextrapolation of dose between species. For instance, in one non-limitingexample, HED may be derived from rhesus monkey dose by multiplying therhesus monkey dose by about 0.324.

In some embodiments of any of the above aspects of the invention, theCXCR2 agonist (e.g., Gro-β or Gro-β T, such as unmodified Gro-β or Gro-βT) is administered intravenously to the donor.

In some embodiments of any of the above aspects of the invention, theCXCR4 antagonist is a compound represented by formula (I)

Z-linker-Z′  (I)

or a pharmaceutically acceptable salt thereof, wherein Z is:

-   -   (i) a cyclic polyamine containing from 9 to 32 ring members,        wherein from 2 to 8 of the ring members are nitrogen atoms        separated from one another by 2 or more carbon atoms; or    -   (ii) an amine represented by formula (IA)

wherein A includes a monocyclic or bicyclic fused ring system includingat least one nitrogen atom and B is H or a substituent of from 1 to 20atoms;

-   -   and wherein Z′ is:    -   (i) a cyclic polyamine containing from 9 to 32 ring members,        wherein from 2 to 8 of the ring members are nitrogen atoms        separated from one another by 2 or more carbon atoms;    -   (ii) an amine represented by formula (IB)

wherein A′ includes a monocyclic or bicyclic fused ring system includingat least one nitrogen atom and B′ is H or a substituent of from 1 to 20atoms; or

-   -   (iii) a substituent represented by formula (IC)

—N(R)—(CR₂)_(n)—X  (IC)

wherein each R is independently H or C₁-C₆ alkyl, n is 1 or 2, and X isan aryl or heteroaryl group or a mercaptan;

wherein the linker is a bond, optionally substituted C₁-C₆ alkylene,optionally substituted C₁-C₆ heteroalkylene, optionally substitutedC₂-C₆ alkenylene, optionally substituted C₂-C₆ heteroalkenylene,optionally substituted C₂-C₆ alkynylene, optionally substituted C₂-C₆heteroalkynylene, optionally substituted cycloalkylene, optionallysubstituted heterocycloalkylene, optionally substituted arylene, oroptionally substituted heteroarylene.

In some embodiments, Z and Z′ are each independently a cyclic polyaminecontaining from 9 to 32 ring members, of which from 2 to 8 are nitrogenatoms separated from one another by 2 or more carbon atoms. Z and Z′ maybe identical substituents. In some embodiments, Z and/or Z′ is a cyclicpolyamine including from 10 to 24 ring members, such as a cyclicpolyamine including 14 ring members. In some embodiments, Z includes 4nitrogen atoms. Z and/or Z′ may be, for example,1,4,8,11-tetraazocyclotetradecane.

In some embodiments, the linker is represented by formula (ID)

wherein ring D is an optionally substituted aryl group, an optionallysubstituted heteroaryl group, an optionally substituted cycloalkylgroup, or an optionally substituted heterocycloalkyl group; and

X and Y are each independently optionally substituted C₁-C₆ alkylene,optionally substituted C₁-C₆ heteroalkylene, optionally substitutedC₂-C₆ alkenylene, optionally substituted C₂-C₆ heteroalkenylene,optionally substituted C₂-C₆ alkynylene, or optionally substituted C₂-C₆heteroalkynylene.

In some embodiments, the linker is represented by formula (IE)

wherein ring D is an optionally substituted aryl group, an optionallysubstituted heteroaryl group, an optionally substituted cycloalkylgroup, or an optionally substituted heterocycloalkyl group; and

X and Y are each independently optionally substituted C₁-C₆ alkylene,optionally substituted C₁-C₆ heteroalkylene, optionally substitutedC₂-C₆ alkenylene, optionally substituted C₂-C₆ heteroalkenylene,optionally substituted C₂-C₆ alkynylene, or optionally substituted C₂-C₆heteroalkynylene.

In some embodiments, X and Y are each independently optionallysubstituted C₁-C₆ alkylene. In some embodiments, X and Y are identicalsubstituents, such as identical alkylene substituents (e.g., methylene,ethylene, propylene, or butylene substituents).

In some embodiments, the CXCR4 antagonist is plerixafor or apharmaceutically acceptable salt thereof. In some embodiments, the CXCR4antagonist (e.g., plerixafor or a pharmaceutically acceptable saltthereof) is administered subcutaneously to the donor. In someembodiments, the CXCR4 antagonist (e.g., plerixafor or apharmaceutically acceptable salt thereof) is administered to the donorat a dose of from about 50 μg/kg to about 500 μg/kg, such as a dose ofabout 50 μg/kg, 55 μg/kg, 60 μg/kg, 65 μg/kg, 70 μg/kg, 75 μg/kg, 80μg/kg, 85 μg/kg, 90 μg/kg, 95 μg/kg, 100 μg/kg, 105 μg/kg, 110 μg/kg,115 μg/kg, 120 μg/kg, 125 μg/kg, 130 μg/kg, 135 μg/kg, 140 μg/kg, 145μg/kg, 150 μg/kg, 155 μg/kg, 160 μg/kg, 165 μg/kg, 170 μg/kg, 175 μg/kg,180 μg/kg, 185 μg/kg, 190 μg/kg, 195 μg/kg, 200 μg/kg, 205 μg/kg, 210μg/kg, 215 μg/kg, 220 μg/kg, 225 μg/kg, 230 μg/kg, 235 μg/kg, 240 μg/kg,245 μg/kg, 250 μg/kg, 255 μg/kg, 260 μg/kg, 265 μg/kg, 270 μg/kg, 275μg/kg, 280 μg/kg, 285 μg/kg, 290 μg/kg, 295 μg/kg, 300 μg/kg, 305 μg/kg,310 μg/kg, 315 μg/kg, 320 μg/kg, 325 μg/kg, 330 μg/kg, 335 μg/kg, 340μg/kg, 345 μg/kg, 350 μg/kg, 355 μg/kg, 360 μg/kg, 365 μg/kg, 370 μg/kg,375 μg/kg, 380 μg/kg, 385 μg/kg, 390 μg/kg, 395 μg/kg, 400 μg/kg, 405μg/kg, 410 μg/kg, 415 μg/kg, 420 μg/kg, 425 μg/kg, 430 μg/kg, 435 μg/kg,440 μg/kg, 445 μg/kg, 450 μg/kg, 455 μg/kg, 460 μg/kg, 465 μg/kg, 470μg/kg, 475 μg/kg, 480 μg/kg, 485 μg/kg, 490 μg/kg, 495 μg/kg, or 500μg/kg. In some embodiments, the CXCR4 antagonist (e.g., plerixafor or apharmaceutically acceptable salt thereof) is administered to the donorat a dose of from about 200 μg/kg to about 300 μg/kg, such as a dose ofabout 240 μg/kg.

For example, in some embodiments, the CXCR4 antagonist (e.g., plerixaforor a pharmaceutically acceptable salt thereof) is administered to thedonor at a dose of from about 50 μg/kg per day to about 500 μg/kg perday, such as a dose of about 50 μg/kg per day, 55 μg/kg per day, 60μg/kg per day, 65 μg/kg per day, 70 μg/kg per day, 75 μg/kg per day, 80μg/kg per day, 85 μg/kg per day, 90 μg/kg per day, 95 μg/kg per day, 100μg/kg per day, 105 μg/kg per day, 110 μg/kg per day, 115 μg/kg per day,120 μg/kg per day, 125 μg/kg per day, 130 μg/kg per day, 135 μg/kg perday, 140 μg/kg per day, 145 μg/kg per day, 150 μg/kg per day, 155 μg/kgper day, 160 μg/kg per day, 165 μg/kg per day, 170 μg/kg per day, 175μg/kg per day, 180 μg/kg per day, 185 μg/kg per day, 190 μg/kg per day,195 μg/kg per day, 200 μg/kg per day, 205 μg/kg per day, 210 μg/kg perday, 215 μg/kg per day, 220 μg/kg per day, 225 μg/kg per day, 230 μg/kgper day, 235 μg/kg per day, 240 μg/kg per day, 245 μg/kg per day, 250μg/kg per day, 255 μg/kg per day, 260 μg/kg per day, 265 μg/kg per day,270 μg/kg per day, 275 μg/kg per day, 280 μg/kg per day, 285 μg/kg perday, 290 μg/kg per day, 295 μg/kg per day, 300 μg/kg per day, 305 μg/kgper day, 310 μg/kg per day, 315 μg/kg per day, 320 μg/kg per day, 325μg/kg per day, 330 μg/kg per day, 335 μg/kg per day, 340 μg/kg per day,345 μg/kg per day, 350 μg/kg per day, 355 μg/kg per day, 360 μg/kg perday, 365 μg/kg per day, 370 μg/kg per day, 375 μg/kg per day, 380 μg/kgper day, 385 μg/kg per day, 390 μg/kg per day, 395 μg/kg per day, 400μs/kg per day, 405 μg/kg per day, 410 μg/kg per day, 415 μg/kg per day,420 μg/kg per day, 425 μg/kg per day, 430 μg/kg per day, 435 μg/kg perday, 440 μg/kg per day, 445 μg/kg per day, 450 μg/kg per day, 455 μg/kgper day, 460 μg/kg per day, 465 μg/kg per day, 470 μg/kg per day, 475μg/kg per day, 480 μg/kg per day, 485 μg/kg per day, 490 μg/kg per day,495 μg/kg per day, or 500 μg/kg per day. In some embodiments, the CXCR4antagonist (e.g., plerixafor or a pharmaceutically acceptable saltthereof) is administered to the donor at a dose of from about 200 μg/kgper day to about 300 μg/kg per day, such as a dose of about 240 μg/kgper day. In some embodiments, the CXCR4 antagonist may be administeredas a single dose. In other embodiments, the CXCR4 antagonist may beadministered as two or more doses.

In some embodiments of any of the above aspects of the invention, theCXCR2 agonist and the CXCR4 antagonist are administered to the donorconcurrently. In some embodiments, the CXCR4 antagonist is administeredto the donor prior to administration of the CXCR2 agonist. In someembodiments, the CXCR4 antagonist may be administered to the donor fromabout 1 minute to about 180 minutes prior to administration of the CXCR2agonist, such as from about 15 minutes to about 180 minutes, about 30minutes to about 180 minutes, about 40 minutes to about 160 minutes,about 50 minutes to about 150 minutes, about 60 minutes to about 140minutes, about 70 minutes to about 130 minutes, about 60 minutes toabout 120 minutes, about 70 minutes to about 110 minutes, or about 80minutes to about 100 minutes (e.g., about 30 minutes, about 35 minutes,about 40 minutes, about 45 minutes, about 50 minutes, about 55 minutes,about 60 minutes, about 65 minutes, about 70 minutes, about 75 minutes,about 80 minutes, about 85 minutes, about 90 minutes, about 95 minutes,about 100 minutes, about 105 minutes, about 110 minutes, about 115minutes, about 120 minutes, about 125 minutes, about 130 minutes, about135 minutes, about 140 minutes, about 145 minutes, about 150 minutes,about 155 minutes, about 160 minutes, about 165 minutes, about 170minutes, about 175 minutes, or about 180 minutes prior to administrationof the CXCR2 agonist). In some embodiments, the CXCR4 antagonist isadministered to the donor from about 30 minutes to about 60 minutesprior to administration of the CXCR2 agonist (e.g., about 30 minutes,about 35 minutes, about 40 minutes, about 45 minutes, about 50 minutes,about 55 minutes, or about 60 minutes prior to administration of theCXCR2 agonist). In some embodiments, the CXCR4 antagonist may beadministered to the donor about 45 minutes prior to administration ofthe CXCR2 agonist.

In a further aspect, the invention features a pharmaceutical compositionincluding a population of hematopoietic stem cells or progeny thereofisolated from a mammalian donor (e.g., a human donor), wherein the ratioof CD34⁺ cells to leukocytes in the population is from about 0.0008 toabout 0.0021. In some embodiments, the ratio of CD34⁺ cells toleukocytes may be about 0.00080, 0.00081, 0.00082, 0.00083, 0.00084,0.00085, 0.00086, 0.00087, 0.00088, 0.00089, 0.00090, 0.00091, 0.00092,0.00093, 0.00094, 0.00095, 0.00096, 0.00097, 0.00098, 0.00099, 0.00100,0.00101, 0.00102, 0.00103, 0.00104, 0.00105, 0.00106, 0.00107, 0.00108,0.00109, 0.00110, 0.00111, 0.00112, 0.00113, 0.00114, 0.00115, 0.00116,0.00117, 0.00118, 0.00119, 0.00120, 0.00121, 0.00122, 0.00123, 0.00124,0.00125, 0.00126, 0.00127, 0.00128, 0.00129, 0.00130, 0.00131, 0.00132,0.00133, 0.00134, 0.00135, 0.00136, 0.00137, 0.00138, 0.00139, 0.00140,0.00141, 0.00142, 0.00143, 0.00144, 0.00145, 0.00146, 0.00147, 0.00148,0.00149, 0.00150, 0.00151, 0.00152, 0.00153, 0.00154, 0.00155, 0.00156,0.00157, 0.00158, 0.00159, 0.00160, 0.00161, 0.00162, 0.00163, 0.00164,0.00165, 0.00166, 0.00167, 0.00168, 0.00169, 0.00170, 0.00171, 0.00172,0.00173, 0.00174, 0.00175, 0.00176, 0.00178, 0.00179, 0.00180, 0.00181,0.00182, 0.00183, 0.00184, 0.00185, 0.00186, 0.00187, 0.00188, 0.00189,0.00190, 0.00191, 0.00192, 0.00193, 0.00194, 0.00195, 0.00196, 0.00197,0.00198, 0.00199, 0.00200, 0.00201, 0.00202, 0.00203, 0.00204, 0.00205,0.00206, 0.00207, 0.00208, 0.00209, 0.00210, 0.00211, 0.00212, 0.00213,0.00214, 0.00215, 0.00216, 0.00217, 0.00218, 0.00219, 0.00220, 0.00221,0.00222, 0.00223, 0.00224, or 0.00225. In some embodiments, the ratio ofCD34⁺ cells to leukocytes is from about 0.0010 to about 0.0018, such asa ratio of CD34⁺ cells to leukocytes of about 0.00100, 0.00101, 0.00102,0.00103, 0.00104, 0.00105, 0.00106, 0.00107, 0.00108, 0.00109, 0.00110,0.00111, 0.00112, 0.00113, 0.00114, 0.00115, 0.00116, 0.00117, 0.00118,0.00119, 0.00120, 0.00121, 0.00122, 0.00123, 0.00124, 0.00125, 0.00126,0.00127, 0.00128, 0.00129, 0.00130, 0.00131, 0.00132, 0.00133, 0.00134,0.00135, 0.00136, 0.00137, 0.00138, 0.00139, 0.00140, 0.00141, 0.00142,0.00143, 0.00144, 0.00145, 0.00146, 0.00147, 0.00148, 0.00149, 0.00150,0.00151, 0.00152, 0.00153, 0.00154, 0.00155, 0.00156, 0.00157, 0.00158,0.00159, 0.00160, 0.00161, 0.00162, 0.00163, 0.00164, 0.00165, 0.00166,0.00167, 0.00168, 0.00169, 0.00170, 0.00171, 0.00172, 0.00173, 0.00174,0.00175, 0.00176, 0.00178, 0.00179, or 0.00180. In some embodiments, theratio of CD34⁺ cells to leukocytes is about 0.0014.

In an additional aspect, the invention features a pharmaceuticalcomposition including a population of hematopoietic stem cells orprogeny thereof isolated from a mammalian donor (e.g., a human donor),wherein the ratio of CD34⁺ cells to neutrophils in the population isfrom about 0.0018 to about 0.0058. In some embodiments, the ratio ofCD34⁺ cells to neutrophils may be about 0.00180, 0.00181, 0.00182,0.00183, 0.00184, 0.00185, 0.00186, 0.00187, 0.00188, 0.00189, 0.00190,0.00191, 0.00192, 0.00193, 0.00194, 0.00195, 0.00196, 0.00197, 0.00198,0.00199, 0.00200, 0.00201, 0.00202, 0.00203, 0.00204, 0.00205, 0.00206,0.00207, 0.00208, 0.00209, 0.00210, 0.00211, 0.00212, 0.00213, 0.00214,0.00215, 0.00216, 0.00217, 0.00218, 0.00219, 0.00220, 0.00221, 0.00222,0.00223, 0.00224, 0.00225, 0.00226, 0.00227, 0.00228, 0.00229, 0.00230,0.00231, 0.00232, 0.00233, 0.00234, 0.00235, 0.00236, 0.00237, 0.00238,0.00239, 0.00240, 0.00241, 0.00242, 0.00243, 0.00244, 0.00245, 0.00246,0.00247, 0.00248, 0.00249, 0.00250, 0.00251, 0.00252, 0.00253, 0.00254,0.00255, 0.00256, 0.00257, 0.00258, 0.00259, 0.00260, 0.00261, 0.00262,0.00263, 0.00264, 0.00265, 0.00266, 0.00267, 0.00268, 0.00269, 0.00270,0.00271, 0.00272, 0.00273, 0.00274, 0.00275, 0.00276, 0.00277, 0.00278,0.00279, 0.00280, 0.00281, 0.00282, 0.00283, 0.00284, 0.00285, 0.00286,0.00287, 0.00288, 0.00289, 0.00290, 0.00291, 0.00292, 0.00293, 0.00294,0.00295, 0.00296, 0.00297, 0.00298, 0.00299, 0.00300, 0.00300, 0.00301,0.00302, 0.00303, 0.00304, 0.00305, 0.00306, 0.00307, 0.00308, 0.00309,0.00310, 0.00311, 0.00312, 0.00313, 0.00314, 0.00315, 0.00316, 0.00317,0.00318, 0.00319, 0.00320, 0.00321, 0.00322, 0.00323, 0.00324, 0.00325,0.00326, 0.00327, 0.00328, 0.00329, 0.00330, 0.00331, 0.00332, 0.00333,0.00334, 0.00335, 0.00336, 0.00337, 0.00338, 0.00339, 0.00340, 0.00341,0.00342, 0.00343, 0.00344, 0.00345, 0.00346, 0.00347, 0.00348, 0.00349,0.00350, 0.00351, 0.00352, 0.00353, 0.00354, 0.00355, 0.00356, 0.00357,0.00358, 0.00359, 0.00360, 0.00361, 0.00362, 0.00363, 0.00364, 0.00365,0.00366, 0.00367, 0.00368, 0.00369, 0.00370, 0.00371, 0.00372, 0.00373,0.00374, 0.00375, 0.00376, 0.00377, 0.00378, 0.00379, 0.00380, 0.00381,0.00382, 0.00383, 0.00384, 0.00385, 0.00386, 0.00387, 0.00388, 0.00389,0.00390, 0.00391, 0.00392, 0.00393, 0.00394, 0.00395, 0.00396, 0.00397,0.00398, 0.00399, 0.00400, 0.00401, 0.00402, 0.00403, 0.00404, 0.00405,0.00406, 0.00407, 0.00408, 0.00409, 0.00410, 0.00411, 0.00412, 0.00413,0.00414, 0.00415, 0.00416, 0.00417, 0.00418, 0.00419, 0.00420, 0.00421,0.00422, 0.00423, 0.00424, 0.00425, 0.00426, 0.00427, 0.00428, 0.00429,0.00430, 0.00431, 0.00432, 0.00433, 0.00434, 0.00435, 0.00436, 0.00437,0.00438, 0.00439, 0.00440, 0.00441, 0.00442, 0.00443, 0.00444, 0.00445,0.00446, 0.00447, 0.00448, 0.00449, 0.00450, 0.00451, 0.00452, 0.00453,0.00454, 0.00455, 0.00456, 0.00457, 0.00458, 0.00459, 0.00460, 0.00461,0.00462, 0.00463, 0.00464, 0.00465, 0.00466, 0.00467, 0.00468, 0.00469,0.00470, 0.00471, 0.00472, 0.00473, 0.00474, 0.00475, 0.00476, 0.00477,0.00478, 0.00479, 0.00480, 0.00481, 0.00482, 0.00483, 0.00484, 0.00485,0.00486, 0.00487, 0.00488, 0.00489, 0.00490, 0.00491, 0.00492, 0.00493,0.00494, 0.00495, 0.00496, 0.00497, 0.00498, 0.00499, 0.00500, 0.00501,0.00502, 0.00503, 0.00504, 0.00505, 0.00506, 0.00507, 0.00508, 0.00509,0.00510, 0.00511, 0.00512, 0.00513, 0.00514, 0.00515, 0.00516, 0.00517,0.00518, 0.00519, 0.00520, 0.00521, 0.00522, 0.00523, 0.00524, 0.00525,0.00526, 0.00527, 0.00528, 0.00529, 0.00530, 0.00531, 0.00532, 0.00533,0.00534, 0.00535, 0.00536, 0.00537, 0.00538, 0.00539, 0.00540, 0.00541,0.00542, 0.00543, 0.00544, 0.00545, 0.00546, 0.00547, 0.00548, 0.00549,0.00550, 0.00551, 0.00552, 0.00553, 0.00554, 0.00555, 0.00556, 0.00557,0.00558, 0.00559, 0.00560, 0.00561, 0.00562, 0.00563, 0.00564, 0.00565,0.00566, 0.00567, 0.00568, 0.00569, 0.00570, 0.00571, 0.00572, 0.00573,0.00574, 0.00575, 0.00576, 0.00577, 0.00578, 0.00579, or 0.00580. Insome embodiments, the ratio of CD34⁺ cells to neutrophils is from about0.0026 to about 0.0046, such as a ratio of CD34⁺ cells to neutrophils ofabout 0.00260, 0.00261, 0.00262, 0.00263, 0.00264, 0.00265, 0.00266,0.00267, 0.00268, 0.00269, 0.00270, 0.00271, 0.00272, 0.00273, 0.00274,0.00275, 0.00276, 0.00277, 0.00278, 0.00279, 0.00280, 0.00281, 0.00282,0.00283, 0.00284, 0.00285, 0.00286, 0.00287, 0.00288, 0.00289, 0.00290,0.00291, 0.00292, 0.00293, 0.00294, 0.00295, 0.00296, 0.00297, 0.00298,0.00299, 0.00300, 0.00300, 0.00301, 0.00302, 0.00303, 0.00304, 0.00305,0.00306, 0.00307, 0.00308, 0.00309, 0.00310, 0.00311, 0.00312, 0.00313,0.00314, 0.00315, 0.00316, 0.00317, 0.00318, 0.00319, 0.00320, 0.00321,0.00322, 0.00323, 0.00324, 0.00325, 0.00326, 0.00327, 0.00328, 0.00329,0.00330, 0.00331, 0.00332, 0.00333, 0.00334, 0.00335, 0.00336, 0.00337,0.00338, 0.00339, 0.00340, 0.00341, 0.00342, 0.00343, 0.00344, 0.00345,0.00346, 0.00347, 0.00348, 0.00349, 0.00350, 0.00351, 0.00352, 0.00353,0.00354, 0.00355, 0.00356, 0.00357, 0.00358, 0.00359, 0.00360, 0.00361,0.00362, 0.00363, 0.00364, 0.00365, 0.00366, 0.00367, 0.00368, 0.00369,0.00370, 0.00371, 0.00372, 0.00373, 0.00374, 0.00375, 0.00376, 0.00377,0.00378, 0.00379, 0.00380, 0.00381, 0.00382, 0.00383, 0.00384, 0.00385,0.00386, 0.00387, 0.00388, 0.00389, 0.00390, 0.00391, 0.00392, 0.00393,0.00394, 0.00395, 0.00396, 0.00397, 0.00398, 0.00399, 0.00400, 0.00401,0.00402, 0.00403, 0.00404, 0.00405, 0.00406, 0.00407, 0.00408, 0.00409,0.00410, 0.00411, 0.00412, 0.00413, 0.00414, 0.00415, 0.00416, 0.00417,0.00418, 0.00419, 0.00420, 0.00421, 0.00422, 0.00423, 0.00424, 0.00425,0.00426, 0.00427, 0.00428, 0.00429, 0.00430, 0.00431, 0.00432, 0.00433,0.00434, 0.00435, 0.00436, 0.00437, 0.00438, 0.00439, 0.00440, 0.00441,0.00442, 0.00443, 0.00444, 0.00445, 0.00446, 0.00447, 0.00448, 0.00449,0.00450, 0.00451, 0.00452, 0.00453, 0.00454, 0.00455, 0.00456, 0.00457,0.00458, 0.00459, or 0.00460. In some embodiments, the ratio of CD34⁺cells to neutrophils is about 0.0036.

In another aspect, the invention features a pharmaceutical compositionincluding a population of hematopoietic stem cells or progeny thereofisolated from a mammalian donor (e.g., a human donor), wherein the ratioof CD34⁺ cells to lymphocytes in the population is from about 0.0021 toabout 0.0094. In some embodiments, the ratio of CD34⁺ cells tolymphocytes may be about 0.00210, 0.00211, 0.00212, 0.00213, 0.00214,0.00215, 0.00216, 0.00217, 0.00218, 0.00219, 0.00220, 0.00221, 0.00222,0.00223, 0.00224, 0.00225, 0.00226, 0.00227, 0.00228, 0.00229, 0.00230,0.00231, 0.00232, 0.00233, 0.00234, 0.00235, 0.00236, 0.00237, 0.00238,0.00239, 0.00240, 0.00241, 0.00242, 0.00243, 0.00244, 0.00245, 0.00246,0.00247, 0.00248, 0.00249, 0.00250, 0.00251, 0.00252, 0.00253, 0.00254,0.00255, 0.00256, 0.00257, 0.00258, 0.00259, 0.00260, 0.00261, 0.00262,0.00263, 0.00264, 0.00265, 0.00266, 0.00267, 0.00268, 0.00269, 0.00270,0.00271, 0.00272, 0.00273, 0.00274, 0.00275, 0.00276, 0.00277, 0.00278,0.00279, 0.00280, 0.00281, 0.00282, 0.00283, 0.00284, 0.00285, 0.00286,0.00287, 0.00288, 0.00289, 0.00290, 0.00291, 0.00292, 0.00293, 0.00294,0.00295, 0.00296, 0.00297, 0.00298, 0.00299, 0.00300, 0.00300, 0.00301,0.00302, 0.00303, 0.00304, 0.00305, 0.00306, 0.00307, 0.00308, 0.00309,0.00310, 0.00311, 0.00312, 0.00313, 0.00314, 0.00315, 0.00316, 0.00317,0.00318, 0.00319, 0.00320, 0.00321, 0.00322, 0.00323, 0.00324, 0.00325,0.00326, 0.00327, 0.00328, 0.00329, 0.00330, 0.00331, 0.00332, 0.00333,0.00334, 0.00335, 0.00336, 0.00337, 0.00338, 0.00339, 0.00340, 0.00341,0.00342, 0.00343, 0.00344, 0.00345, 0.00346, 0.00347, 0.00348, 0.00349,0.00350, 0.00351, 0.00352, 0.00353, 0.00354, 0.00355, 0.00356, 0.00357,0.00358, 0.00359, 0.00360, 0.00361, 0.00362, 0.00363, 0.00364, 0.00365,0.00366, 0.00367, 0.00368, 0.00369, 0.00370, 0.00371, 0.00372, 0.00373,0.00374, 0.00375, 0.00376, 0.00377, 0.00378, 0.00379, 0.00380, 0.00381,0.00382, 0.00383, 0.00384, 0.00385, 0.00386, 0.00387, 0.00388, 0.00389,0.00390, 0.00391, 0.00392, 0.00393, 0.00394, 0.00395, 0.00396, 0.00397,0.00398, 0.00399, 0.00400, 0.00401, 0.00402, 0.00403, 0.00404, 0.00405,0.00406, 0.00407, 0.00408, 0.00409, 0.00410, 0.00411, 0.00412, 0.00413,0.00414, 0.00415, 0.00416, 0.00417, 0.00418, 0.00419, 0.00420, 0.00421,0.00422, 0.00423, 0.00424, 0.00425, 0.00426, 0.00427, 0.00428, 0.00429,0.00430, 0.00431, 0.00432, 0.00433, 0.00434, 0.00435, 0.00436, 0.00437,0.00438, 0.00439, 0.00440, 0.00441, 0.00442, 0.00443, 0.00444, 0.00445,0.00446, 0.00447, 0.00448, 0.00449, 0.00450, 0.00451, 0.00452, 0.00453,0.00454, 0.00455, 0.00456, 0.00457, 0.00458, 0.00459, 0.00460, 0.00461,0.00462, 0.00463, 0.00464, 0.00465, 0.00466, 0.00467, 0.00468, 0.00469,0.00470, 0.00471, 0.00472, 0.00473, 0.00474, 0.00475, 0.00476, 0.00477,0.00478, 0.00479, 0.00480, 0.00481, 0.00482, 0.00483, 0.00484, 0.00485,0.00486, 0.00487, 0.00488, 0.00489, 0.00490, 0.00491, 0.00492, 0.00493,0.00494, 0.00495, 0.00496, 0.00497, 0.00498, 0.00499, 0.00500, 0.00501,0.00502, 0.00503, 0.00504, 0.00505, 0.00506, 0.00507, 0.00508, 0.00509,0.00510, 0.00511, 0.00512, 0.00513, 0.00514, 0.00515, 0.00516, 0.00517,0.00518, 0.00519, 0.00520, 0.00521, 0.00522, 0.00523, 0.00524, 0.00525,0.00526, 0.00527, 0.00528, 0.00529, 0.00530, 0.00531, 0.00532, 0.00533,0.00534, 0.00535, 0.00536, 0.00537, 0.00538, 0.00539, 0.00540, 0.00541,0.00542, 0.00543, 0.00544, 0.00545, 0.00546, 0.00547, 0.00548, 0.00549,0.00550, 0.00551, 0.00552, 0.00553, 0.00554, 0.00555, 0.00556, 0.00557,0.00558, 0.00559, 0.00560, 0.00561, 0.00562, 0.00563, 0.00564, 0.00565,0.00566, 0.00567, 0.00568, 0.00569, 0.00570, 0.00571, 0.00572, 0.00573,0.00574, 0.00575, 0.00576, 0.00577, 0.00578, 0.00579, 0.00580, 0.00581,0.00582, 0.00583, 0.00584, 0.00585, 0.00586, 0.00587, 0.00588, 0.00589,0.00590, 0.00591, 0.00592, 0.00593, 0.00594, 0.00595, 0.00596, 0.00597,0.00598, 0.00599, 0.00600, 0.00601, 0.00602, 0.00603, 0.00604, 0.00605,0.00606, 0.00607, 0.00608, 0.00609, 0.00610, 0.00611, 0.00612, 0.00613,0.00614, 0.00615, 0.00616, 0.00617, 0.00618, 0.00619, 0.00620, 0.00621,0.00622, 0.00623, 0.00624, 0.00625, 0.00626, 0.00627, 0.00628, 0.00629,0.00630, 0.00631, 0.00632, 0.00633, 0.00634, 0.00635, 0.00636, 0.00637,0.00638, 0.00639, 0.00640, 0.00641, 0.00642, 0.00643, 0.00644, 0.00645,0.00646, 0.00647, 0.00648, 0.00649, 0.00650, 0.00651, 0.00652, 0.00653,0.00654, 0.00655, 0.00656, 0.00657, 0.00658, 0.00659, 0.00660, 0.00661,0.00662, 0.00663, 0.00664, 0.00665, 0.00666, 0.00667, 0.00668, 0.00669,0.00670, 0.00671, 0.00672, 0.00673, 0.00674, 0.00675, 0.00676, 0.00677,0.00678, 0.00679, 0.00680, 0.00681, 0.00682, 0.00683, 0.00684, 0.00685,0.00686, 0.00687, 0.00688, 0.00689, 0.00690, 0.00691, 0.00692, 0.00693,0.00694, 0.00695, 0.00696, 0.00697, 0.00698, 0.00699, 0.00700, 0.00701,0.00702, 0.00703, 0.00704, 0.00705, 0.00706, 0.00707, 0.00708, 0.00709,0.00710, 0.00711, 0.00712, 0.00713, 0.00714, 0.00715, 0.00716, 0.00717,0.00718, 0.00719, 0.00720, 0.00721, 0.00722, 0.00723, 0.00724, 0.00725,0.00726, 0.00727, 0.00728, 0.00729, 0.00730, 0.00731, 0.00732, 0.00733,0.00734, 0.00735, 0.00736, 0.00737, 0.00738, 0.00739, 0.00740, 0.00741,0.00742, 0.00743, 0.00744, 0.00745, 0.00746, 0.00747, 0.00748, 0.00749,0.00750, 0.00751, 0.00752, 0.00753, 0.00754, 0.00755, 0.00756, 0.00757,0.00758, 0.00759, 0.00760, 0.00761, 0.00762, 0.00763, 0.00764, 0.00765,0.00766, 0.00767, 0.00768, 0.00769, 0.00770, 0.00771, 0.00772, 0.00773,0.00774, 0.00775, 0.00776, 0.00777, 0.00778, 0.00779, 0.00780, 0.00781,0.00782, 0.00783, 0.00784, 0.00785, 0.00786, 0.00787, 0.00788, 0.00789,0.00790, 0.00791, 0.00792, 0.00793, 0.00794, 0.00795, 0.00796, 0.00797,0.00798, 0.00799, 0.00800, 0.00801, 0.00802, 0.00803, 0.00804, 0.00805,0.00806, 0.00807, 0.00808, 0.00809, 0.00810, 0.00811, 0.00812, 0.00813,0.00814, 0.00815, 0.00816, 0.00817, 0.00818, 0.00819, 0.00820, 0.00821,0.00822, 0.00823, 0.00824, 0.00825, 0.00826, 0.00827, 0.00828, 0.00829,0.00830, 0.00831, 0.00832, 0.00833, 0.00834, 0.00835, 0.00836, 0.00837,0.00838, 0.00839, 0.00840, 0.00841, 0.00842, 0.00843, 0.00844, 0.00845,0.00846, 0.00847, 0.00848, 0.00849, 0.00850, 0.00851, 0.00852, 0.00853,0.00854, 0.00855, 0.00856, 0.00857, 0.00858, 0.00859, 0.00860, 0.00861,0.00862, 0.00863, 0.00864, 0.00865, 0.00866, 0.00867, 0.00868, 0.00869,0.00870, 0.00871, 0.00872, 0.00873, 0.00874, 0.00875, 0.00876, 0.00877,0.00878, 0.00879, 0.00880, 0.00881, 0.00882, 0.00883, 0.00884, 0.00885,0.00886, 0.00887, 0.00888, 0.00889, 0.00890, 0.00891, 0.00892, 0.00893,0.00894, 0.00895, 0.00896, 0.00897, 0.00898, 0.00899, 0.00900, 0.00901,0.00902, 0.00903, 0.00904, 0.00905, 0.00906, 0.00907, 0.00908, 0.00909,0.00910, 0.00911, 0.00912, 0.00913, 0.00914, 0.00915, 0.00916, 0.00917,0.00918, 0.00919, 0.00920, 0.00921, 0.00922, 0.00923, 0.00924, 0.00925,0.00926, 0.00927, 0.00928, 0.00929, 0.00930, 0.00931, 0.00932, 0.00933,0.00934, 0.00935, 0.00936, 0.00937, 0.00938, 0.00939, or 0.00940. Insome embodiments, the ratio of CD34⁺ cells to lymphocytes is from about0.0025 to about 0.0035, such as a ratio of CD34⁺ cells to lymphocytes ofabout 0.00250, 0.00251, 0.00252, 0.00253, 0.00254, 0.00255, 0.00256,0.00257, 0.00258, 0.00259, 0.00260, 0.00261, 0.00262, 0.00263, 0.00264,0.00265, 0.00266, 0.00267, 0.00268, 0.00269, 0.00270, 0.00271, 0.00272,0.00273, 0.00274, 0.00275, 0.00276, 0.00277, 0.00278, 0.00279, 0.00280,0.00281, 0.00282, 0.00283, 0.00284, 0.00285, 0.00286, 0.00287, 0.00288,0.00289, 0.00290, 0.00291, 0.00292, 0.00293, 0.00294, 0.00295, 0.00296,0.00297, 0.00298, 0.00299, 0.00300, 0.00300, 0.00301, 0.00302, 0.00303,0.00304, 0.00305, 0.00306, 0.00307, 0.00308, 0.00309, 0.00310, 0.00311,0.00312, 0.00313, 0.00314, 0.00315, 0.00316, 0.00317, 0.00318, 0.00319,0.00320, 0.00321, 0.00322, 0.00323, 0.00324, 0.00325, 0.00326, 0.00327,0.00328, 0.00329, 0.00330, 0.00331, 0.00332, 0.00333, 0.00334, 0.00335,0.00336, 0.00337, 0.00338, 0.00339, 0.00340, 0.00341, 0.00342, 0.00343,0.00344, 0.00345, 0.00346, 0.00347, 0.00348, 0.00349, or 0.00350. Insome embodiments, the ratio of CD34⁺ cells to lymphocytes is about0.0031.

In a further aspect, the invention features a pharmaceutical compositionincluding a population of hematopoietic stem cells or progeny thereofisolated from a mammalian donor (e.g., a human donor), wherein the ratioof CD34⁺ cells to monocytes in the population is from about 0.0071 toabout 0.0174. In some embodiments, the ratio of CD34⁺ cells to monocytesmay be about 0.00710, 0.00711, 0.00712, 0.00713, 0.00714, 0.00715,0.00716, 0.00717, 0.00718, 0.00719, 0.00720, 0.00721, 0.00722, 0.00723,0.00724, 0.00725, 0.00726, 0.00727, 0.00728, 0.00729, 0.00730, 0.00731,0.00732, 0.00733, 0.00734, 0.00735, 0.00736, 0.00737, 0.00738, 0.00739,0.00740, 0.00741, 0.00742, 0.00743, 0.00744, 0.00745, 0.00746, 0.00747,0.00748, 0.00749, 0.00750, 0.00751, 0.00752, 0.00753, 0.00754, 0.00755,0.00756, 0.00757, 0.00758, 0.00759, 0.00760, 0.00761, 0.00762, 0.00763,0.00764, 0.00765, 0.00766, 0.00767, 0.00768, 0.00769, 0.00770, 0.00771,0.00772, 0.00773, 0.00774, 0.00775, 0.00776, 0.00777, 0.00778, 0.00779,0.00780, 0.00781, 0.00782, 0.00783, 0.00784, 0.00785, 0.00786, 0.00787,0.00788, 0.00789, 0.00790, 0.00791, 0.00792, 0.00793, 0.00794, 0.00795,0.00796, 0.00797, 0.00798, 0.00799, 0.00800, 0.00801, 0.00802, 0.00803,0.00804, 0.00805, 0.00806, 0.00807, 0.00808, 0.00809, 0.00810, 0.00811,0.00812, 0.00813, 0.00814, 0.00815, 0.00816, 0.00817, 0.00818, 0.00819,0.00820, 0.00821, 0.00822, 0.00823, 0.00824, 0.00825, 0.00826, 0.00827,0.00828, 0.00829, 0.00830, 0.00831, 0.00832, 0.00833, 0.00834, 0.00835,0.00836, 0.00837, 0.00838, 0.00839, 0.00840, 0.00841, 0.00842, 0.00843,0.00844, 0.00845, 0.00846, 0.00847, 0.00848, 0.00849, 0.00850, 0.00851,0.00852, 0.00853, 0.00854, 0.00855, 0.00856, 0.00857, 0.00858, 0.00859,0.00860, 0.00861, 0.00862, 0.00863, 0.00864, 0.00865, 0.00866, 0.00867,0.00868, 0.00869, 0.00870, 0.00871, 0.00872, 0.00873, 0.00874, 0.00875,0.00876, 0.00877, 0.00878, 0.00879, 0.00880, 0.00881, 0.00882, 0.00883,0.00884, 0.00885, 0.00886, 0.00887, 0.00888, 0.00889, 0.00890, 0.00891,0.00892, 0.00893, 0.00894, 0.00895, 0.00896, 0.00897, 0.00898, 0.00899,0.00900, 0.00901, 0.00902, 0.00903, 0.00904, 0.00905, 0.00906, 0.00907,0.00908, 0.00909, 0.00910, 0.00911, 0.00912, 0.00913, 0.00914, 0.00915,0.00916, 0.00917, 0.00918, 0.00919, 0.00920, 0.00921, 0.00922, 0.00923,0.00924, 0.00925, 0.00926, 0.00927, 0.00928, 0.00929, 0.00930, 0.00931,0.00932, 0.00933, 0.00934, 0.00935, 0.00936, 0.00937, 0.00938, 0.00939,0.00940, 0.00941, 0.00942, 0.00943, 0.00944, 0.00945, 0.00946, 0.00947,0.00948, 0.00949, 0.00950, 0.00951, 0.00952, 0.00953, 0.00954, 0.00955,0.00956, 0.00957, 0.00958, 0.00959, 0.00960, 0.00961, 0.00962, 0.00963,0.00964, 0.00965, 0.00966, 0.00967, 0.00968, 0.00969, 0.00970, 0.00971,0.00972, 0.00973, 0.00974, 0.00975, 0.00976, 0.00977, 0.00978, 0.00979,0.00980, 0.00981, 0.00982, 0.00983, 0.00984, 0.00985, 0.00986, 0.00987,0.00988, 0.00989, 0.00990, 0.00991, 0.00992, 0.00993, 0.00994, 0.00995,0.00996, 0.00997, 0.00998, 0.00999, 0.0100, 0.0101, 0.0103, 0.0104,0.0105, 0.0106, 0.0107, 0.0108, 0.0109, 0.0110, 0.0111, 0.0112, 0.0113,0.0114, 0.0115, 0.0116, 0.0117, 0.0118, 0.0119, 0.0120, 0.0121, 0.0122,0.0123, 0.0124, 0.0125, 0.0126, 0.0127, 0.0128, 0.0129, 0.0130, 0.0131,0.0132, 0.0133, 0.0134, 0.0135, 0.0136, 0.0137, 0.0138, 0.0139, 0.0140,0.0141, 0.0142, 0.0143, 0.0144, 0.0145, 0.0146, 0.0147, 0.0148, 0.0149,0.0150, 0.0151, 0.0152, 0.0153, 0.0154, 0.0155, 0.0156, 0.0157, 0.0158,0.0159, 0.0160, 0.0161, 0.0162, 0.0163, 0.0164, 0.0165, 0.0166, 0.0167,0.0168, 0.0169, 0.0170, 0.0171, 0.0172, 0.0173, or 0.0174. In someembodiments, the ratio of CD34⁺ cells to monocytes is from about 0.0100to about 0.0140, such as a ratio of CD34⁺ cells to monocytes of about0.0100, 0.0101, 0.0103, 0.0104, 0.0105, 0.0106, 0.0107, 0.0108, 0.0109,0.0110, 0.0111, 0.0112, 0.0113, 0.0114, 0.0115, 0.0116, 0.0117, 0.0118,0.0119, 0.0120, 0.0121, 0.0122, 0.0123, 0.0124, 0.0125, 0.0126, 0.0127,0.0128, 0.0129, 0.0130, 0.0131, 0.0132, 0.0133, 0.0134, 0.0135, 0.0136,0.0137, 0.0138, 0.0139, or 0.0140. In some embodiments, the ratio ofCD34⁺ cells to monocytes is about 0.0118.

In a further aspect, the invention features a pharmaceutical compositionincluding a population of hematopoietic stem cells or progeny thereofisolated from a mammalian donor (e.g., a human donor), wherein thefrequency of CD34⁺ cells in the population is from about 0.051% to about0.140%. In some embodiments, the population of cells may have afrequency of CD34⁺ cells of about 0.051%, 0.052%, 0.053%, 0.054%,0.055%, 0.056%, 0.057%, 0.058%, 0.059%, 0.060%, 0.061%, 0.062%, 0.063%,0.064%, 0.065%, 0.066%, 0.067%, 0.068%, 0.069%, 0.070%, 0.071%, 0.072%,0.073%, 0.074%, 0.075%, 0.076%, 0.077%, 0.078%, 0.079%, 0.080%, 0.081%,0.082%, 0.083%, 0.084%, 0.085%, 0.086%, 0.087%, 0.088%, 0.089%, 0.090%,0.091%, 0.092%, 0.093%, 0.094%, 0.095%, 0.096%, 0.097%, 0.098%, 0.099%,0.100%, 0.101%, 0.102%, 0.103%, 0.104%, 0.105%, 0.106%, 0.107%, 0.108%,0.109%, 0.110%, 0.111%, 0.112%, 0.113%, 0.114%, 0.115%, 0.116%, 0.117%,0.118%, 0.119%, 0.120%, 0.121%, 0.122%, 0.123%, 0.124%, 0.125%, 0.126%,0.127%, 0.128%, 0.129%, 0.130%, 0.131%, 0.132%, 0.133%, 0.134%, 0.135%,0.136%, 0.137%, 0.138%, 0.139%, or 0.140%. In some embodiments, thepopulation of cells has a frequency of CD34⁺ cells of from about 0.080%to about 0.120%, such as a frequency of CD34⁺ cells of about 0.080%,0.081%, 0.082%, 0.083%, 0.084%, 0.085%, 0.086%, 0.087%, 0.088%, 0.089%,0.090%, 0.091%, 0.092%, 0.093%, 0.094%, 0.095%, 0.096%, 0.097%, 0.098%,0.099%, 0.100%, 0.101%, 0.102%, 0.103%, 0.104%, 0.105%, 0.106%, 0.107%,0.108%, 0.109%, 0.110%, 0.111%, 0.112%, 0.113%, 0.114%, 0.115%, 0.116%,0.117%, 0.118%, 0.119%, or 0.120%. In some embodiments, the populationof cells has a frequency of CD34⁺ cells of about 0.097%.

In a further aspect, the invention features a pharmaceutical compositionincluding a population of hematopoietic stem cells or progeny thereofisolated from a mammalian donor (e.g., a human donor), wherein the ratioof CD34⁺ CD90⁺ CD45RA⁻ cells to leukocytes in the population is fromabout 0.0003 to about 0.0016. In some embodiments, the ratio of CD34⁺CD90⁺ CD45RA⁻ cells to leukocytes may be about 0.00030, 0.00031,0.00032, 0.00033, 0.00034, 0.00035, 0.00036, 0.00037, 0.00038, 0.00039,0.00040, 0.00041, 0.00042, 0.00043, 0.00044, 0.00045, 0.00046, 0.00047,0.00048, 0.00049, 0.00050, 0.00051, 0.00052, 0.00053, 0.00054, 0.00055,0.00056, 0.00057, 0.00058, 0.00059, 0.00060, 0.00061, 0.00062, 0.00063,0.00064, 0.00065, 0.00066, 0.00067, 0.00068, 0.00069, 0.00070, 0.00071,0.00072, 0.00073, 0.00074, 0.00075, 0.00076, 0.00077, 0.00078, 0.00079,0.00080, 0.00081, 0.00082, 0.00083, 0.00084, 0.00085, 0.00086, 0.00087,0.00088, 0.00089, 0.00090, 0.00091, 0.00092, 0.00093, 0.00094, 0.00095,0.00096, 0.00097, 0.00098, 0.00099, 0.00100, 0.00101, 0.00102, 0.00103,0.00104, 0.00105, 0.00106, 0.00107, 0.00108, 0.00109, 0.00110, 0.00111,0.00112, 0.00113, 0.00114, 0.00115, 0.00116, 0.00117, 0.00118, 0.00119,0.00120, 0.00121, 0.00122, 0.00123, 0.00124, 0.00125, 0.00126, 0.00127,0.00128, 0.00129, 0.00130, 0.00131, 0.00132, 0.00133, 0.00134, 0.00135,0.00136, 0.00137, 0.00138, 0.00139, 0.00140, 0.00141, 0.00142, 0.00143,0.00144, 0.00145, 0.00146, 0.00147, 0.00148, 0.00149, 0.00150, 0.00151,0.00152, 0.00153, 0.00154, 0.00155, 0.00156, 0.00157, 0.00158, 0.00159,or 0.00160. In some embodiments, the ratio of CD34⁺ CD90⁺ CD45RA⁻ cellsto leukocytes is from about 0.0006 to about 0.0012, such as a ratio ofCD34⁺ CD90⁺ CD45RA⁻ cells to leukocytes of about 0.00060, 0.00061,0.00062, 0.00063, 0.00064, 0.00065, 0.00066, 0.00067, 0.00068, 0.00069,0.00070, 0.00071, 0.00072, 0.00073, 0.00074, 0.00075, 0.00076, 0.00077,0.00078, 0.00079, 0.00080, 0.00081, 0.00082, 0.00083, 0.00084, 0.00085,0.00086, 0.00087, 0.00088, 0.00089, 0.00090, 0.00091, 0.00092, 0.00093,0.00094, 0.00095, 0.00096, 0.00097, 0.00098, 0.00099, 0.00100, 0.00101,0.00102, 0.00103, 0.00104, 0.00105, 0.00106, 0.00107, 0.00108, 0.00109,0.00110, 0.00111, 0.00112, 0.00113, 0.00114, 0.00115, 0.00116, 0.00117,0.00118, 0.00119, or 0.00120. In some embodiments, the ratio of CD34⁺CD90⁺ CD45RA⁻ cells to leukocytes is about 0.0009.

In an additional aspect, the invention features a pharmaceuticalcomposition including a population of hematopoietic stem cells orprogeny thereof isolated from a mammalian donor (e.g., a human donor),wherein the ratio of CD34⁺ CD90⁺ CD45RA⁻ cells to neutrophils in thepopulation is from about 0.0007 to about 0.0043. In some embodiments,the ratio of CD34⁺ CD90⁺ CD45RA⁻ cells to neutrophils may be about0.00070, 0.00071, 0.00072, 0.00073, 0.00074, 0.00075, 0.00076, 0.00077,0.00078, 0.00079, 0.00080, 0.00081, 0.00082, 0.00083, 0.00084, 0.00085,0.00086, 0.00087, 0.00088, 0.00089, 0.00090, 0.00091, 0.00092, 0.00093,0.00094, 0.00095, 0.00096, 0.00097, 0.00098, 0.00099, 0.00100, 0.00101,0.00102, 0.00103, 0.00104, 0.00105, 0.00106, 0.00107, 0.00108, 0.00109,0.00110, 0.00111, 0.00112, 0.00113, 0.00114, 0.00115, 0.00116, 0.00117,0.00118, 0.00119, 0.00120, 0.00121, 0.00122, 0.00123, 0.00124, 0.00125,0.00126, 0.00127, 0.00128, 0.00129, 0.00130, 0.00131, 0.00132, 0.00133,0.00134, 0.00135, 0.00136, 0.00137, 0.00138, 0.00139, 0.00140, 0.00141,0.00142, 0.00143, 0.00144, 0.00145, 0.00146, 0.00147, 0.00148, 0.00149,0.00150, 0.00151, 0.00152, 0.00153, 0.00154, 0.00155, 0.00156, 0.00157,0.00158, 0.00159, 0.00160, 0.00161, 0.00162, 0.00163, 0.00164, 0.00165,0.00166, 0.00167, 0.00168, 0.00169, 0.00170, 0.00171, 0.00172, 0.00173,0.00174, 0.00175, 0.00176, 0.00177, 0.00178, 0.00179, 0.00180, 0.00181,0.00182, 0.00183, 0.00184, 0.00185, 0.00186, 0.00187, 0.00188, 0.00189,0.00190, 0.00191, 0.00192, 0.00193, 0.00194, 0.00195, 0.00196, 0.00197,0.00198, 0.00199, 0.00200, 0.00201, 0.00202, 0.00203, 0.00204, 0.00205,0.00206, 0.00207, 0.00208, 0.00209, 0.00210, 0.00211, 0.00212, 0.00213,0.00214, 0.00215, 0.00216, 0.00217, 0.00218, 0.00219, 0.00220, 0.00221,0.00222, 0.00223, 0.00224, 0.00225, 0.00226, 0.00227, 0.00228, 0.00229,0.00230, 0.00231, 0.00232, 0.00233, 0.00234, 0.00235, 0.00236, 0.00237,0.00238, 0.00239, 0.00240, 0.00241, 0.00242, 0.00243, 0.00244, 0.00245,0.00246, 0.00247, 0.00248, 0.00249, 0.00250, 0.00251, 0.00252, 0.00253,0.00254, 0.00255, 0.00256, 0.00257, 0.00258, 0.00259, 0.00260, 0.00261,0.00262, 0.00263, 0.00264, 0.00265, 0.00266, 0.00267, 0.00268, 0.00269,0.00270, 0.00271, 0.00272, 0.00273, 0.00274, 0.00275, 0.00276, 0.00277,0.00278, 0.00279, 0.00280, 0.00281, 0.00282, 0.00283, 0.00284, 0.00285,0.00286, 0.00287, 0.00288, 0.00289, 0.00290, 0.00291, 0.00292, 0.00293,0.00294, 0.00295, 0.00296, 0.00297, 0.00298, 0.00299, 0.00300, 0.00300,0.00301, 0.00302, 0.00303, 0.00304, 0.00305, 0.00306, 0.00307, 0.00308,0.00309, 0.00310, 0.00311, 0.00312, 0.00313, 0.00314, 0.00315, 0.00316,0.00317, 0.00318, 0.00319, 0.00320, 0.00321, 0.00322, 0.00323, 0.00324,0.00325, 0.00326, 0.00327, 0.00328, 0.00329, 0.00330, 0.00331, 0.00332,0.00333, 0.00334, 0.00335, 0.00336, 0.00337, 0.00338, 0.00339, 0.00340,0.00341, 0.00342, 0.00343, 0.00344, 0.00345, 0.00346, 0.00347, 0.00348,0.00349, 0.00350, 0.00351, 0.00352, 0.00353, 0.00354, 0.00355, 0.00356,0.00357, 0.00358, 0.00359, 0.00360, 0.00361, 0.00362, 0.00363, 0.00364,0.00365, 0.00366, 0.00367, 0.00368, 0.00369, 0.00370, 0.00371, 0.00372,0.00373, 0.00374, 0.00375, 0.00376, 0.00377, 0.00378, 0.00379, 0.00380,0.00381, 0.00382, 0.00383, 0.00384, 0.00385, 0.00386, 0.00387, 0.00388,0.00389, 0.00390, 0.00391, 0.00392, 0.00393, 0.00394, 0.00395, 0.00396,0.00397, 0.00398, 0.00399, 0.00400, 0.00401, 0.00402, 0.00403, 0.00404,0.00405, 0.00406, 0.00407, 0.00408, 0.00409, 0.00410, 0.00411, 0.00412,0.00413, 0.00414, 0.00415, 0.00416, 0.00417, 0.00418, 0.00419, 0.00420,0.00421, 0.00422, 0.00423, 0.00424, 0.00425, 0.00426, 0.00427, 0.00428,0.00429, or 0.00430. In some embodiments, the ratio of CD34⁺ CD90⁺CD45RA⁻ cells to neutrophils is from about 0.0014 to about 0.0034, suchas a ratio of CD34⁺ CD90⁺ CD45RA⁻ cells to neutrophils of about 0.00140,0.00141, 0.00142, 0.00143, 0.00144, 0.00145, 0.00146, 0.00147, 0.00148,0.00149, 0.00150, 0.00151, 0.00152, 0.00153, 0.00154, 0.00155, 0.00156,0.00157, 0.00158, 0.00159, 0.00160, 0.00161, 0.00162, 0.00163, 0.00164,0.00165, 0.00166, 0.00167, 0.00168, 0.00169, 0.00170, 0.00171, 0.00172,0.00173, 0.00174, 0.00175, 0.00176, 0.00177, 0.00178, 0.00179, 0.00180,0.00181, 0.00182, 0.00183, 0.00184, 0.00185, 0.00186, 0.00187, 0.00188,0.00189, 0.00190, 0.00191, 0.00192, 0.00193, 0.00194, 0.00195, 0.00196,0.00197, 0.00198, 0.00199, 0.00200, 0.00201, 0.00202, 0.00203, 0.00204,0.00205, 0.00206, 0.00207, 0.00208, 0.00209, 0.00210, 0.00211, 0.00212,0.00213, 0.00214, 0.00215, 0.00216, 0.00217, 0.00218, 0.00219, 0.00220,0.00221, 0.00222, 0.00223, 0.00224, 0.00225, 0.00226, 0.00227, 0.00228,0.00229, 0.00230, 0.00231, 0.00232, 0.00233, 0.00234, 0.00235, 0.00236,0.00237, 0.00238, 0.00239, 0.00240, 0.00241, 0.00242, 0.00243, 0.00244,0.00245, 0.00246, 0.00247, 0.00248, 0.00249, 0.00250, 0.00251, 0.00252,0.00253, 0.00254, 0.00255, 0.00256, 0.00257, 0.00258, 0.00259, 0.00260,0.00261, 0.00262, 0.00263, 0.00264, 0.00265, 0.00266, 0.00267, 0.00268,0.00269, 0.00270, 0.00271, 0.00272, 0.00273, 0.00274, 0.00275, 0.00276,0.00277, 0.00278, 0.00279, 0.00280, 0.00281, 0.00282, 0.00283, 0.00284,0.00285, 0.00286, 0.00287, 0.00288, 0.00289, 0.00290, 0.00291, 0.00292,0.00293, 0.00294, 0.00295, 0.00296, 0.00297, 0.00298, 0.00299, 0.00300,0.00300, 0.00301, 0.00302, 0.00303, 0.00304, 0.00305, 0.00306, 0.00307,0.00308, 0.00309, 0.00310, 0.00311, 0.00312, 0.00313, 0.00314, 0.00315,0.00316, 0.00317, 0.00318, 0.00319, 0.00320, 0.00321, 0.00322, 0.00323,0.00324, 0.00325, 0.00326, 0.00327, 0.00328, 0.00329, 0.00330, 0.00331,0.00332, 0.00333, 0.00334, 0.00335, 0.00336, 0.00337, 0.00338, 0.00339,or 0.00340. In some embodiments, the ratio of CD34⁺ CD90⁺ CD45RA⁻ cellsto neutrophils is about 0.0024.

In another aspect, the invention features a pharmaceutical compositionincluding a population of hematopoietic stem cells or progeny thereofisolated from a mammalian donor (e.g., a human donor), wherein the ratioof CD34⁺ CD90⁺ CD45RA⁻ cells to lymphocytes in the population is fromabout 0.0008 to about 0.0069. In some embodiments, the ratio of CD34⁺CD90⁺ CD45RA⁻ cells to lymphocytes may be about 0.00080, 0.00081,0.00082, 0.00083, 0.00084, 0.00085, 0.00086, 0.00087, 0.00088, 0.00089,0.00090, 0.00091, 0.00092, 0.00093, 0.00094, 0.00095, 0.00096, 0.00097,0.00098, 0.00099, 0.00100, 0.00101, 0.00102, 0.00103, 0.00104, 0.00105,0.00106, 0.00107, 0.00108, 0.00109, 0.00110, 0.00111, 0.00112, 0.00113,0.00114, 0.00115, 0.00116, 0.00117, 0.00118, 0.00119, 0.00120, 0.00121,0.00122, 0.00123, 0.00124, 0.00125, 0.00126, 0.00127, 0.00128, 0.00129,0.00130, 0.00131, 0.00132, 0.00133, 0.00134, 0.00135, 0.00136, 0.00137,0.00138, 0.00139, 0.00140, 0.00141, 0.00142, 0.00143, 0.00144, 0.00145,0.00146, 0.00147, 0.00148, 0.00149, 0.00150, 0.00151, 0.00152, 0.00153,0.00154, 0.00155, 0.00156, 0.00157, 0.00158, 0.00159, 0.00160, 0.00161,0.00162, 0.00163, 0.00164, 0.00165, 0.00166, 0.00167, 0.00168, 0.00169,0.00170, 0.00171, 0.00172, 0.00173, 0.00174, 0.00175, 0.00176, 0.00178,0.00179, 0.00180, 0.00181, 0.00182, 0.00183, 0.00184, 0.00185, 0.00186,0.00187, 0.00188, 0.00189, 0.00190, 0.00191, 0.00192, 0.00193, 0.00194,0.00195, 0.00196, 0.00197, 0.00198, 0.00199, 0.00200, 0.00201, 0.00202,0.00203, 0.00204, 0.00205, 0.00206, 0.00207, 0.00208, 0.00209, 0.00210,0.00211, 0.00212, 0.00213, 0.00214, 0.00215, 0.00216, 0.00217, 0.00218,0.00219, 0.00220, 0.00221, 0.00222, 0.00223, 0.00224, 0.00225, 0.00226,0.00227, 0.00228, 0.00229, 0.00230, 0.00231, 0.00232, 0.00233, 0.00234,0.00235, 0.00236, 0.00237, 0.00238, 0.00239, 0.00240, 0.00241, 0.00242,0.00243, 0.00244, 0.00245, 0.00246, 0.00247, 0.00248, 0.00249, 0.00250,0.00251, 0.00252, 0.00253, 0.00254, 0.00255, 0.00256, 0.00257, 0.00258,0.00259, 0.00260, 0.00261, 0.00262, 0.00263, 0.00264, 0.00265, 0.00266,0.00267, 0.00268, 0.00269, 0.00270, 0.00271, 0.00272, 0.00273, 0.00274,0.00275, 0.00276, 0.00278, 0.00279, 0.00280, 0.00281, 0.00282, 0.00283,0.00284, 0.00285, 0.00286, 0.00287, 0.00288, 0.00289, 0.00290, 0.00291,0.00292, 0.00293, 0.00294, 0.00295, 0.00296, 0.00297, 0.00298, 0.00299,0.00300, 0.00301, 0.00302, 0.00303, 0.00304, 0.00305, 0.00306, 0.00307,0.00308, 0.00309, 0.00310, 0.00311, 0.00312, 0.00313, 0.00314, 0.00315,0.00316, 0.00317, 0.00318, 0.00319, 0.00320, 0.00321, 0.00322, 0.00323,0.00324, 0.00325, 0.00326, 0.00327, 0.00328, 0.00329, 0.00330, 0.00331,0.00332, 0.00333, 0.00334, 0.00335, 0.00336, 0.00337, 0.00338, 0.00339,0.00340, 0.00341, 0.00342, 0.00343, 0.00344, 0.00345, 0.00346, 0.00347,0.00348, 0.00349, 0.00350, 0.00351, 0.00352, 0.00353, 0.00354, 0.00355,0.00356, 0.00357, 0.00358, 0.00359, 0.00360, 0.00361, 0.00362, 0.00363,0.00364, 0.00365, 0.00366, 0.00367, 0.00368, 0.00369, 0.00370, 0.00371,0.00372, 0.00373, 0.00374, 0.00375, 0.00376, 0.00378, 0.00379, 0.00380,0.00381, 0.00382, 0.00383, 0.00384, 0.00385, 0.00386, 0.00387, 0.00388,0.00389, 0.00390, 0.00391, 0.00392, 0.00393, 0.00394, 0.00395, 0.00396,0.00397, 0.00398, 0.00399, 0.00401, 0.00402, 0.00403, 0.00404, 0.00405,0.00406, 0.00407, 0.00408, 0.00409, 0.00410, 0.00411, 0.00412, 0.00413,0.00414, 0.00415, 0.00416, 0.00417, 0.00418, 0.00419, 0.00420, 0.00421,0.00422, 0.00423, 0.00424, 0.00425, 0.00426, 0.00427, 0.00428, 0.00429,0.00430, 0.00431, 0.00432, 0.00433, 0.00434, 0.00435, 0.00436, 0.00437,0.00438, 0.00439, 0.00440, 0.00441, 0.00442, 0.00443, 0.00444, 0.00445,0.00446, 0.00447, 0.00448, 0.00449, 0.00450, 0.00451, 0.00452, 0.00453,0.00454, 0.00455, 0.00456, 0.00457, 0.00458, 0.00459, 0.00460, 0.00461,0.00462, 0.00463, 0.00464, 0.00465, 0.00466, 0.00467, 0.00468, 0.00469,0.00470, 0.00471, 0.00472, 0.00473, 0.00474, 0.00475, 0.00476, 0.00478,0.00479, 0.00480, 0.00481, 0.00482, 0.00483, 0.00484, 0.00485, 0.00486,0.00487, 0.00488, 0.00489, 0.00490, 0.00491, 0.00492, 0.00493, 0.00494,0.00495, 0.00496, 0.00497, 0.00498, 0.00499, 0.00500, 0.00501, 0.00502,0.00503, 0.00504, 0.00505, 0.00506, 0.00507, 0.00508, 0.00509, 0.00510,0.00511, 0.00512, 0.00513, 0.00514, 0.00515, 0.00516, 0.00517, 0.00518,0.00519, 0.00520, 0.00521, 0.00522, 0.00523, 0.00524, 0.00525, 0.00526,0.00527, 0.00528, 0.00529, 0.00530, 0.00531, 0.00532, 0.00533, 0.00534,0.00535, 0.00536, 0.00537, 0.00538, 0.00539, 0.00540, 0.00541, 0.00542,0.00543, 0.00544, 0.00545, 0.00546, 0.00547, 0.00548, 0.00549, 0.00550,0.00551, 0.00552, 0.00553, 0.00554, 0.00555, 0.00556, 0.00557, 0.00558,0.00559, 0.00560, 0.00561, 0.00562, 0.00563, 0.00564, 0.00565, 0.00566,0.00567, 0.00568, 0.00569, 0.00570, 0.00571, 0.00572, 0.00573, 0.00574,0.00575, 0.00576, 0.00578, 0.00579, 0.00580, 0.00581, 0.00582, 0.00583,0.00584, 0.00585, 0.00586, 0.00587, 0.00588, 0.00589, 0.00590, 0.00591,0.00592, 0.00593, 0.00594, 0.00595, 0.00596, 0.00597, 0.00598, 0.00599,0.00600, 0.00601, 0.00602, 0.00603, 0.00604, 0.00605, 0.00606, 0.00607,0.00608, 0.00609, 0.00610, 0.00611, 0.00612, 0.00613, 0.00614, 0.00615,0.00616, 0.00617, 0.00618, 0.00619, 0.00620, 0.00621, 0.00622, 0.00623,0.00624, 0.00625, 0.00626, 0.00627, 0.00628, 0.00629, 0.00630, 0.00631,0.00632, 0.00633, 0.00634, 0.00635, 0.00636, 0.00637, 0.00638, 0.00639,0.00640, 0.00641, 0.00642, 0.00643, 0.00644, 0.00645, 0.00646, 0.00647,0.00648, 0.00649, 0.00650, 0.00651, 0.00652, 0.00653, 0.00654, 0.00655,0.00656, 0.00657, 0.00658, 0.00659, 0.00660, 0.00661, 0.00662, 0.00663,0.00664, 0.00665, 0.00666, 0.00667, 0.00668, 0.00669, 0.00670, 0.00671,0.00672, 0.00673, 0.00674, 0.00675, 0.00676, 0.00678, 0.00679, 0.00680,0.00681, 0.00682, 0.00683, 0.00684, 0.00685, 0.00686, 0.00687, 0.00688,0.00689, or 0.00690. In some embodiments, the ratio of CD34⁺ CD90⁺CD45RA⁻ cells to lymphocytes is from about 0.0011 to about 0.0031, suchas a ratio of CD34⁺ CD90⁺ CD45RA⁻ cells to lymphocytes of about 0.00110,0.00111, 0.00112, 0.00113, 0.00114, 0.00115, 0.00116, 0.00117, 0.00118,0.00119, 0.00120, 0.00121, 0.00122, 0.00123, 0.00124, 0.00125, 0.00126,0.00127, 0.00128, 0.00129, 0.00130, 0.00131, 0.00132, 0.00133, 0.00134,0.00135, 0.00136, 0.00137, 0.00138, 0.00139, 0.00140, 0.00141, 0.00142,0.00143, 0.00144, 0.00145, 0.00146, 0.00147, 0.00148, 0.00149, 0.00150,0.00151, 0.00152, 0.00153, 0.00154, 0.00155, 0.00156, 0.00157, 0.00158,0.00159, 0.00160, 0.00161, 0.00162, 0.00163, 0.00164, 0.00165, 0.00166,0.00167, 0.00168, 0.00169, 0.00170, 0.00171, 0.00172, 0.00173, 0.00174,0.00175, 0.00176, 0.00178, 0.00179, 0.00180, 0.00181, 0.00182, 0.00183,0.00184, 0.00185, 0.00186, 0.00187, 0.00188, 0.00189, 0.00190, 0.00191,0.00192, 0.00193, 0.00194, 0.00195, 0.00196, 0.00197, 0.00198, 0.00199,0.00200, 0.00201, 0.00202, 0.00203, 0.00204, 0.00205, 0.00206, 0.00207,0.00208, 0.00209, 0.00210, 0.00211, 0.00212, 0.00213, 0.00214, 0.00215,0.00216, 0.00217, 0.00218, 0.00219, 0.00220, 0.00221, 0.00222, 0.00223,0.00224, 0.00225, 0.00226, 0.00227, 0.00228, 0.00229, 0.00230, 0.00231,0.00232, 0.00233, 0.00234, 0.00235, 0.00236, 0.00237, 0.00238, 0.00239,0.00240, 0.00241, 0.00242, 0.00243, 0.00244, 0.00245, 0.00246, 0.00247,0.00248, 0.00249, 0.00250, 0.00251, 0.00252, 0.00253, 0.00254, 0.00255,0.00256, 0.00257, 0.00258, 0.00259, 0.00260, 0.00261, 0.00262, 0.00263,0.00264, 0.00265, 0.00266, 0.00267, 0.00268, 0.00269, 0.00270, 0.00271,0.00272, 0.00273, 0.00274, 0.00275, 0.00276, 0.00278, 0.00279, 0.00280,0.00281, 0.00282, 0.00283, 0.00284, 0.00285, 0.00286, 0.00287, 0.00288,0.00289, 0.00290, 0.00291, 0.00292, 0.00293, 0.00294, 0.00295, 0.00296,0.00297, 0.00298, 0.00299, 0.00300, 0.00301, 0.00302, 0.00303, 0.00304,0.00305, 0.00306, 0.00307, 0.00308, 0.00309, or 0.00310. In someembodiments, the ratio of CD34⁺ CD90⁺ CD45RA⁻-cells to lymphocytes isabout 0.0021.

In a further aspect, the invention features a pharmaceutical compositionincluding a population of hematopoietic stem cells or progeny thereofisolated from a mammalian donor (e.g., a human donor), wherein the ratioof CD34⁺ CD90⁺ CD45RA⁻ cells to monocytes in the population is fromabout 0.0028 to about 0.0130. In some embodiments, the ratio of CD34⁺CD90⁺ CD45RA⁻ cells to monocytes may be about 0.00280, 0.00281, 0.00282,0.00283, 0.00284, 0.00285, 0.00286, 0.00287, 0.00288, 0.00289, 0.00290,0.00291, 0.00292, 0.00293, 0.00294, 0.00295, 0.00296, 0.00297, 0.00298,0.00299, 0.00300, 0.00301, 0.00302, 0.00303, 0.00304, 0.00305, 0.00306,0.00307, 0.00308, 0.00309, 0.00310, 0.00311, 0.00312, 0.00313, 0.00314,0.00315, 0.00316, 0.00317, 0.00318, 0.00319, 0.00320, 0.00321, 0.00322,0.00323, 0.00324, 0.00325, 0.00326, 0.00327, 0.00328, 0.00329, 0.00330,0.00331, 0.00332, 0.00333, 0.00334, 0.00335, 0.00336, 0.00337, 0.00338,0.00339, 0.00340, 0.00341, 0.00342, 0.00343, 0.00344, 0.00345, 0.00346,0.00347, 0.00348, 0.00349, 0.00350, 0.00351, 0.00352, 0.00353, 0.00354,0.00355, 0.00356, 0.00357, 0.00358, 0.00359, 0.00360, 0.00361, 0.00362,0.00363, 0.00364, 0.00365, 0.00366, 0.00367, 0.00368, 0.00369, 0.00370,0.00371, 0.00372, 0.00373, 0.00374, 0.00375, 0.00376, 0.00378, 0.00379,0.00380, 0.00381, 0.00382, 0.00383, 0.00384, 0.00385, 0.00386, 0.00387,0.00388, 0.00389, 0.00390, 0.00391, 0.00392, 0.00393, 0.00394, 0.00395,0.00396, 0.00397, 0.00398, 0.00399, 0.00401, 0.00402, 0.00403, 0.00404,0.00405, 0.00406, 0.00407, 0.00408, 0.00409, 0.00410, 0.00411, 0.00412,0.00413, 0.00414, 0.00415, 0.00416, 0.00417, 0.00418, 0.00419, 0.00420,0.00421, 0.00422, 0.00423, 0.00424, 0.00425, 0.00426, 0.00427, 0.00428,0.00429, 0.00430, 0.00431, 0.00432, 0.00433, 0.00434, 0.00435, 0.00436,0.00437, 0.00438, 0.00439, 0.00440, 0.00441, 0.00442, 0.00443, 0.00414,0.00445, 0.00446, 0.00447, 0.00448, 0.00449, 0.00450, 0.00451, 0.00452,0.00453, 0.00454, 0.00455, 0.00456, 0.00457, 0.00458, 0.00459, 0.00460,0.00461, 0.00462, 0.00463, 0.00464, 0.00465, 0.00466, 0.00467, 0.00468,0.00469, 0.00470, 0.00471, 0.00472, 0.00473, 0.00474, 0.00475, 0.00476,0.00478, 0.00479, 0.00480, 0.00481, 0.00482, 0.00483, 0.00484, 0.00485,0.00486, 0.00487, 0.00488, 0.00489, 0.00490, 0.00491, 0.00492, 0.00493,0.00494, 0.00495, 0.00496, 0.00497, 0.00498, 0.00499, 0.00500, 0.00501,0.00502, 0.00503, 0.00504, 0.00505, 0.00506, 0.00507, 0.00508, 0.00509,0.00510, 0.00511, 0.00512, 0.00513, 0.00514, 0.00515, 0.00516, 0.00517,0.00518, 0.00519, 0.00520, 0.00521, 0.00522, 0.00523, 0.00524, 0.00525,0.00526, 0.00527, 0.00528, 0.00529, 0.00530, 0.00531, 0.00532, 0.00533,0.00534, 0.00535, 0.00536, 0.00537, 0.00538, 0.00539, 0.00540, 0.00541,0.00542, 0.00543, 0.00544, 0.00545, 0.00546, 0.00547, 0.00548, 0.00549,0.00550, 0.00551, 0.00552, 0.00553, 0.00554, 0.00555, 0.00556, 0.00557,0.00558, 0.00559, 0.00560, 0.00561, 0.00562, 0.00563, 0.00564, 0.00565,0.00566, 0.00567, 0.00568, 0.00569, 0.00570, 0.00571, 0.00572, 0.00573,0.00574, 0.00575, 0.00576, 0.00578, 0.00579, 0.00580, 0.00581, 0.00582,0.00583, 0.00584, 0.00585, 0.00586, 0.00587, 0.00588, 0.00589, 0.00590,0.00591, 0.00592, 0.00593, 0.00594, 0.00595, 0.00596, 0.00597, 0.00598,0.00599, 0.00600, 0.00601, 0.00602, 0.00603, 0.00604, 0.00605, 0.00606,0.00607, 0.00608, 0.00609, 0.00610, 0.00611, 0.00612, 0.00613, 0.00614,0.00615, 0.00616, 0.00617, 0.00618, 0.00619, 0.00620, 0.00621, 0.00622,0.00623, 0.00624, 0.00625, 0.00626, 0.00627, 0.00628, 0.00629, 0.00630,0.00631, 0.00632, 0.00633, 0.00634, 0.00635, 0.00636, 0.00637, 0.00638,0.00639, 0.00640, 0.00641, 0.00642, 0.00643, 0.00644, 0.00645, 0.00646,0.00647, 0.00648, 0.00649, 0.00650, 0.00651, 0.00652, 0.00653, 0.00654,0.00655, 0.00656, 0.00657, 0.00658, 0.00659, 0.00660, 0.00661, 0.00662,0.00663, 0.00664, 0.00665, 0.00666, 0.00667, 0.00668, 0.00669, 0.00670,0.00671, 0.00672, 0.00673, 0.00674, 0.00675, 0.00676, 0.00678, 0.00679,0.00680, 0.00681, 0.00682, 0.00683, 0.00684, 0.00685, 0.00686, 0.00687,0.00688, 0.00689, 0.00690, 0.00691, 0.00692, 0.00693, 0.00694, 0.00695,0.00696, 0.00697, 0.00698, 0.00699, 0.00700, 0.00701, 0.00702, 0.00703,0.00704, 0.00705, 0.00706, 0.00707, 0.00708, 0.00709, 0.00710, 0.00711,0.00712, 0.00713, 0.00714, 0.00715, 0.00716, 0.00717, 0.00718, 0.00719,0.00720, 0.00721, 0.00722, 0.00723, 0.00724, 0.00725, 0.00726, 0.00727,0.00728, 0.00729, 0.00730, 0.00731, 0.00732, 0.00733, 0.00734, 0.00735,0.00736, 0.00737, 0.00738, 0.00739, 0.00740, 0.00741, 0.00742, 0.00743,0.00744, 0.00745, 0.00746, 0.00747, 0.00748, 0.00749, 0.00750, 0.00751,0.00752, 0.00753, 0.00754, 0.00755, 0.00756, 0.00757, 0.00758, 0.00759,0.00760, 0.00761, 0.00762, 0.00763, 0.00764, 0.00765, 0.00766, 0.00767,0.00768, 0.00769, 0.00770, 0.00771, 0.00772, 0.00773, 0.00774, 0.00775,0.00776, 0.00777, 0.00778, 0.00779, 0.00780, 0.00781, 0.00782, 0.00783,0.00784, 0.00785, 0.00786, 0.00787, 0.00788, 0.00789, 0.00790, 0.00791,0.00792, 0.00793, 0.00794, 0.00795, 0.00796, 0.00797, 0.00798, 0.00799,0.00800, 0.00801, 0.00802, 0.00803, 0.00804, 0.00805, 0.00806, 0.00807,0.00808, 0.00809, 0.00810, 0.00811, 0.00812, 0.00813, 0.00814, 0.00815,0.00816, 0.00817, 0.00818, 0.00819, 0.00820, 0.00821, 0.00822, 0.00823,0.00824, 0.00825, 0.00826, 0.00827, 0.00828, 0.00829, 0.00830, 0.00831,0.00832, 0.00833, 0.00834, 0.00835, 0.00836, 0.00837, 0.00838, 0.00839,0.00840, 0.00841, 0.00842, 0.00843, 0.00844, 0.00845, 0.00846, 0.00847,0.00848, 0.00849, 0.00850, 0.00851, 0.00852, 0.00853, 0.00854, 0.00855,0.00856, 0.00857, 0.00858, 0.00859, 0.00860, 0.00861, 0.00862, 0.00863,0.00864, 0.00865, 0.00866, 0.00867, 0.00868, 0.00869, 0.00870, 0.00871,0.00872, 0.00873, 0.00874, 0.00875, 0.00876, 0.00877, 0.00878, 0.00879,0.00880, 0.00881, 0.00882, 0.00883, 0.00884, 0.00885, 0.00886, 0.00887,0.00888, 0.00889, 0.00890, 0.00891, 0.00892, 0.00893, 0.00894, 0.00895,0.00896, 0.00897, 0.00898, 0.00899, 0.00900, 0.00901, 0.00902, 0.00903,0.00904, 0.00905, 0.00906, 0.00907, 0.00908, 0.00909, 0.00910, 0.00911,0.00912, 0.00913, 0.00914, 0.00915, 0.00916, 0.00917, 0.00918, 0.00919,0.00920, 0.00921, 0.00922, 0.00923, 0.00924, 0.00925, 0.00926, 0.00927,0.00928, 0.00929, 0.00930, 0.00931, 0.00932, 0.00933, 0.00934, 0.00935,0.00936, 0.00937, 0.00938, 0.00939, 0.00940, 0.00941, 0.00942, 0.00943,0.00944, 0.00945, 0.00946, 0.00947, 0.00948, 0.00949, 0.00950, 0.00951,0.00952, 0.00953, 0.00954, 0.00955, 0.00956, 0.00957, 0.00958, 0.00959,0.00960, 0.00961, 0.00962, 0.00963, 0.00964, 0.00965, 0.00966, 0.00967,0.00968, 0.00969, 0.00970, 0.00971, 0.00972, 0.00973, 0.00974, 0.00975,0.00976, 0.00977, 0.00978, 0.00979, 0.00980, 0.00981, 0.00982, 0.00983,0.00984, 0.00985, 0.00986, 0.00987, 0.00988, 0.00989, 0.00990, 0.00991,0.00992, 0.00993, 0.00994, 0.00995, 0.00996, 0.00997, 0.00998, 0.00999,0.0100, 0.0101, 0.0103, 0.0104, 0.0105, 0.0106, 0.0107, 0.0108, 0.0109,0.0110, 0.0111, 0.0112, 0.0113, 0.0114, 0.0115, 0.0116, 0.0117, 0.0118,0.0119, 0.0120, 0.0121, 0.0122, 0.0123, 0.0124, 0.0125, 0.0126, 0.0127,0.0128, 0.0129, or 0.0130. In some embodiments, the ratio of CD34⁺ CD90⁺CD45RA⁻ cells to monocytes is from about 0.0063 to about 0.0083, such asa ratio of CD34⁺ CD90⁺ CD45RA⁻ cells to monocytes of about 0.00630,0.00631, 0.00632, 0.00633, 0.00634, 0.00635, 0.00636, 0.00637, 0.00638,0.00639, 0.00640, 0.00641, 0.00642, 0.00643, 0.00644, 0.00645, 0.00646,0.00647, 0.00648, 0.00649, 0.00650, 0.00651, 0.00652, 0.00653, 0.00654,0.00655, 0.00656, 0.00657, 0.00658, 0.00659, 0.00660, 0.00661, 0.00662,0.00663, 0.00664, 0.00665, 0.00666, 0.00667, 0.00668, 0.00669, 0.00670,0.00671, 0.00672, 0.00673, 0.00674, 0.00675, 0.00676, 0.00678, 0.00679,0.00680, 0.00681, 0.00682, 0.00683, 0.00684, 0.00685, 0.00686, 0.00687,0.00688, 0.00689, 0.00690, 0.00691, 0.00692, 0.00693, 0.00694, 0.00695,0.00696, 0.00697, 0.00698, 0.00699, 0.00700, 0.00701, 0.00702, 0.00703,0.00704, 0.00705, 0.00706, 0.00707, 0.00708, 0.00709, 0.00710, 0.00711,0.00712, 0.00713, 0.00714, 0.00715, 0.00716, 0.00717, 0.00718, 0.00719,0.00720, 0.00721, 0.00722, 0.00723, 0.00724, 0.00725, 0.00726, 0.00727,0.00728, 0.00729, 0.00730, 0.00731, 0.00732, 0.00733, 0.00734, 0.00735,0.00736, 0.00737, 0.00738, 0.00739, 0.00740, 0.00741, 0.00742, 0.00743,0.00744, 0.00745, 0.00746, 0.00747, 0.00748, 0.00749, 0.00750, 0.00751,0.00752, 0.00753, 0.00754, 0.00755, 0.00756, 0.00757, 0.00758, 0.00759,0.00760, 0.00761, 0.00762, 0.00763, 0.00764, 0.00765, 0.00766, 0.00767,0.00768, 0.00769, 0.00770, 0.00771, 0.00772, 0.00773, 0.00774, 0.00775,0.00776, 0.00777, 0.00778, 0.00779, 0.00780, 0.00781, 0.00782, 0.00783,0.00784, 0.00785, 0.00786, 0.00787, 0.00788, 0.00789, 0.00790, 0.00791,0.00792, 0.00793, 0.00794, 0.00795, 0.00796, 0.00797, 0.00798, 0.00799,0.00800, 0.00801, 0.00802, 0.00803, 0.00804, 0.00805, 0.00806, 0.00807,0.00808, 0.00809, 0.00810, 0.00811, 0.00812, 0.00813, 0.00814, 0.00815,0.00816, 0.00817, 0.00818, 0.00819, 0.00820, 0.00821, 0.00822, 0.00823,0.00824, 0.00825, 0.00826, 0.00827, 0.00828, 0.00829, or 0.00830. Insome embodiments, the ratio of CD34⁺ CD90⁺ CD45RA⁻ cells to monocytes isabout 0.0073.

In a further aspect, the invention features a pharmaceutical compositionincluding a population of hematopoietic stem cells or progeny thereofisolated from a mammalian donor (e.g., a human donor), wherein the ratioof CD34⁺ CD90⁺ CD45RA⁻ cells to CD34⁺ cells in the population is fromabout 0.393 to about 0.745. In some embodiments, the ratio of CD34⁺CD90⁺ CD45RA⁻ cells to CD34⁺ cells may be about 0.393, 0.394, 0.395,0.396, 0.397, 0.398, 0.399, 0.401, 0.402, 0.403, 0.404, 0.405, 0.406,0.407, 0.408, 0.409, 0.410, 0.411, 0.412, 0.413, 0.414, 0.415, 0.416,0.417, 0.418, 0.419, 0.420, 0.421, 0.422, 0.423, 0.424, 0.425, 0.426,0.427, 0.428, 0.429, 0.430, 0.431, 0.432, 0.433, 0.434, 0.435, 0.436,0.437, 0.438, 0.439, 0.440, 0.441, 0.442, 0.443, 0.444, 0.445, 0.446,0.447, 0.448, 0.449, 0.450, 0.451, 0.452, 0.453, 0.454, 0.455, 0.456,0.457, 0.458, 0.459, 0.460, 0.461, 0.462, 0.463, 0.464, 0.465, 0.466,0.467, 0.468, 0.469, 0.470, 0.471, 0.472, 0.473, 0.474, 0.475, 0.476,0.478, 0.479, 0.480, 0.481, 0.482, 0.483, 0.484, 0.485, 0.486, 0.487,0.488, 0.489, 0.490, 0.491, 0.492, 0.493, 0.494, 0.495, 0.496, 0.497,0.498, 0.499, 0.500, 0.501, 0.502, 0.503, 0.504, 0.505, 0.506, 0.507,0.508, 0.509, 0.510, 0.511, 0.512, 0.513, 0.514, 0.515, 0.516, 0.517,0.518, 0.519, 0.520, 0.521, 0.522, 0.523, 0.524, 0.525, 0.526, 0.527,0.528, 0.529, 0.530, 0.531, 0.532, 0.533, 0.534, 0.535, 0.536, 0.537,0.538, 0.539, 0.540, 0.541, 0.542, 0.543, 0.544, 0.545, 0.546, 0.547,0.548, 0.549, 0.550, 0.551, 0.552, 0.553, 0.554, 0.555, 0.556, 0.557,0.558, 0.559, 0.560, 0.561, 0.562, 0.563, 0.564, 0.565, 0.566, 0.567,0.568, 0.569, 0.570, 0.571, 0.572, 0.573, 0.574, 0.575, 0.576, 0.578,0.579, 0.580, 0.581, 0.582, 0.583, 0.584, 0.585, 0.586, 0.587, 0.588,0.589, 0.590, 0.591, 0.592, 0.593, 0.594, 0.595, 0.596, 0.597, 0.598,0.599, 0.600, 0.601, 0.602, 0.603, 0.604, 0.605, 0.606, 0.607, 0.608,0.609, 0.610, 0.611, 0.612, 0.613, 0.614, 0.615, 0.616, 0.617, 0.618,0.619, 0.620, 0.621, 0.622, 0.623, 0.624, 0.625, 0.626, 0.627, 0.628,0.629, 0.630, 0.631, 0.632, 0.633, 0.634, 0.635, 0.636, 0.637, 0.638,0.639, 0.640, 0.641, 0.642, 0.643, 0.644, 0.645, 0.646, 0.647, 0.648,0.649, 0.650, 0.651, 0.652, 0.653, 0.654, 0.655, 0.656, 0.657, 0.658,0.659, 0.660, 0.661, 0.662, 0.663, 0.664, 0.665, 0.666, 0.667, 0.668,0.669, 0.670, 0.671, 0.672, 0.673, 0.674, 0.675, 0.676, 0.678, 0.679,0.680, 0.681, 0.682, 0.683, 0.684, 0.685, 0.686, 0.687, 0.688, 0.689,0.690, 0.691, 0.692, 0.693, 0.694, 0.695, 0.696, 0.697, 0.698, 0.699,0.700, 0.701, 0.702, 0.703, 0.704, 0.705, 0.706, 0.707, 0.708, 0.709,0.710, 0.711, 0.712, 0.713, 0.714, 0.715, 0.716, 0.717, 0.718, 0.719,0.720, 0.721, 0.722, 0.723, 0.724, 0.725, 0.726, 0.727, 0.728, 0.729,0.730, 0.731, 0.732, 0.733, 0.734, 0.735, 0.736, 0.737, 0.738, 0.739,0.740, 0.741, 0.742, 0.743, 0.744, or 0.745. In some embodiments, theratio of CD34⁺ CD90⁺ CD45RA⁻ cells to CD34⁺ cells is from about 0.625 toabout 0.725, such as a ratio of CD34⁺ CD90⁺ CD45RA⁻ cells to CD34⁺ cellsof about 0.625, 0.626, 0.627, 0.628, 0.629, 0.630, 0.631, 0.632, 0.633,0.634, 0.635, 0.636, 0.637, 0.638, 0.639, 0.640, 0.641, 0.642, 0.643,0.644, 0.645, 0.646, 0.647, 0.648, 0.649, 0.650, 0.651, 0.652, 0.653,0.654, 0.655, 0.656, 0.657, 0.658, 0.659, 0.660, 0.661, 0.662, 0.663,0.664, 0.665, 0.666, 0.667, 0.668, 0.669, 0.670, 0.671, 0.672, 0.673,0.674, 0.675, 0.676, 0.678, 0.679, 0.680, 0.681, 0.682, 0.683, 0.684,0.685, 0.686, 0.687, 0.688, 0.689, 0.690, 0.691, 0.692, 0.693, 0.694,0.695, 0.696, 0.697, 0.698, 0.699, 0.700, 0.701, 0.702, 0.703, 0.704,0.705, 0.706, 0.707, 0.708, 0.709, 0.710, 0.711, 0.712, 0.713, 0.714,0.715, 0.716, 0.717, 0.718, 0.719, 0.720, 0.721, 0.722, 0.723, 0.724, or0.725. In some embodiments, the ratio of CD34⁺ CD90⁺ CD45RA⁻ cells toCD34⁺ cells is about 0.676.

In a further aspect, the invention features a pharmaceutical compositionincluding a population of hematopoietic stem cells or progeny thereofisolated from a mammalian donor (e.g., a human donor), wherein thefrequency of CD34⁺ CD90⁺ CD45RA⁻ cells in the population is from about0.020% to about 0.110%. In some embodiments, the population of cells mayhave a frequency of CD34⁺ CD90⁺ CD45RA⁻ cells of about 0.020%, 0.021%,0.022%, 0.023%, 0.024%, 0.025%, 0.026%, 0.027%, 0.028%, 0.029%, 0.030%,0.031%, 0.032%, 0.033%, 0.034%, 0.035%, 0.036%, 0.037%, 0.038%, 0.039%,0.040%, 0.041%, 0.042%, 0.043%, 0.044%, 0.045%, 0.046%, 0.047%, 0.048%,0.049%, 0.050%, 0.051%, 0.052%, 0.053%, 0.054%, 0.055%, 0.056%, 0.057%,0.058%, 0.059%, 0.060%, 0.061%, 0.062%, 0.063%, 0.064%, 0.065%, 0.066%,0.067%, 0.068%, 0.069%, 0.070%, 0.071%, 0.072%, 0.073%, 0.074%, 0.075%,0.076%, 0.077%, 0.078%, 0.079%, 0.080%, 0.081%, 0.082%, 0.083%, 0.084%,0.085%, 0.086%, 0.087%, 0.088%, 0.089%, 0.090%, 0.091%, 0.092%, 0.093%,0.094%, 0.095%, 0.096%, 0.097%, 0.098%, 0.099%, 0.100%, 0.101%, 0.102%,0.103%, 0.104%, 0.105%, 0.106%, 0.107%, 0.108%, 0.109%, or 0.110%. Insome embodiments, the population of cells has a frequency of CD34⁺ CD90⁺CD45RA⁻-cells of from about 0.046% to about 0.086%, such as a frequencyof hematopoietic stem cells of about 0.046%, 0.047%, 0.048%, 0.049%,0.050%, 0.051%, 0.052%, 0.053%, 0.054%, 0.055%, 0.056%, 0.057%, 0.058%,0.059%, 0.060%, 0.061%, 0.062%, 0.063%, 0.064%, 0.065%, 0.066%, 0.067%,0.068%, 0.069%, 0.070%, 0.071%, 0.072%, 0.073%, 0.074%, 0.075%, 0.076%,0.077%, 0.078%, 0.079%, 0.080%, 0.081%, 0.082%, 0.083%, 0.084%, 0.085%,or 0.086%. In some embodiments, the population of cells has a frequencyof CD34⁺ CD90⁺ CD45RA⁻ cells of about 0.066%.

In another aspect, the invention features a method of treating a stemcell disorder in a mammalian patient (e.g., a human patient), the methodincluding mobilizing a population of hematopoietic stem cells in amammalian donor (e.g., a human donor) in accordance with any of theabove-described methods, and infusing a therapeutically effective amountof the hematopoietic stem cells, or progeny thereof, into the patient.

In a further aspect, the invention features a method of treating a stemcell disorder in a mammalian patient (e.g., a human patient), the methodincluding infusing into the patient a therapeutically effective amountof the hematopoietic stem cells mobilized by any of the above-describedmethods, or progeny thereof.

In another aspect, the invention features a method of treating a stemcell disorder in a mammalian patient (e.g., a human patient), the methodincluding administering to the patient any one or more of theabove-described pharmaceutical compositions.

In some embodiments of any of the three preceding aspects, the stem celldisorder is a hemoglobinopathy disorder, such as sickle cell anemia,thalassemia, Fanconi anemia, aplastic anemia, and Wiskott-Aldrichsyndrome. In some embodiments, the stem cell disorder is amyelodysplastic disorder. The stem cell disorder may be animmunodeficiency disorder, such as a congenital immunodeficiency or anacquired immunodeficiency, for example, human immunodeficiency virus oracquired immune deficiency syndrome. In some embodiments, the stem celldisorder is a metabolic disorder, such as a metabolic disorder selectedfrom glycogen storage diseases, mucopolysaccharidoses, Gaucher'sDisease, Hurlers Disease, sphingolipidoses, and metachromaticleukodystrophy.

In some embodiments, the stem cell disorder is cancer. The cancer maybe, for example, leukemia, lymphoma, multiple myeloma, andneuroblastoma. In some embodiments, the cancer is a hematologicalcancer. In some embodiments, the cancer is acute myeloid leukemia, acutelymphoid leukemia, chronic myeloid leukemia, chronic lymphoid leukemia,multiple myeloma, diffuse large B-cell lymphoma, or non-Hodgkin'slymphoma.

In some embodiments, the stem cell disorder is a disorder selected fromthe group consisting of adenosine deaminase deficiency and severecombined immunodeficiency, hyper immunoglobulin M syndrome,Chediak-Higashi disease, hereditary lymphohistiocytosis, osteopetrosis,osteogenesis imperfecta, storage diseases, thalassemia major, systemicsclerosis, systemic lupus erythematosus, multiple sclerosis, andjuvenile rheumatoid arthritis.

In some embodiments, the stem cell disorder is an autoimmune disorder,such as an autoimmune disorder selected from multiple sclerosis, humansystemic lupus, rheumatoid arthritis, inflammatory bowel disease,treating psoriasis, Type 1 diabetes mellitus, acute disseminatedencephalomyelitis, Addison's disease, alopecia universalis, ankylosingspondylitis, antiphospholipid antibody syndrome, aplastic anemia,autoimmune hemolytic anemia, autoimmune hepatitis, autoimmune inner eardisease, autoimmune lymphoproliferative syndrome, autoimmune oophoritis,Balo disease, Behcet's disease, bullous pemphigoid, cardiomyopathy,Chagas' disease, chronic fatigue immune dysfunction syndrome, chronicinflammatory demyelinating polyneuropathy, Crohn's disease, cicatricialpemphigoid, coeliac sprue-dermatitis herpetiformis, cold agglutinindisease, CREST syndrome, Degos disease, discoid lupus, dysautonomia,endometriosis, essential mixed cryoglobulinemia,fibromyalgia-fibromyositis, Goodpasture's syndrome, Grave's disease,Guillain-Barre syndrome, Hashimoto's thyroiditis, Hidradenitissuppurativa, idiopathic and/or acute thrombocytopenic purpura,idiopathic pulmonary fibrosis, IgA neuropathy, interstitial cystitis,juvenile arthritis, Kawasaki's disease, lichen planus, Lyme disease,Meniere disease, mixed connective tissue disease, myasthenia gravis,neuromyotonia, opsoclonus myoclonus syndrome, optic neuritis, Ord'sthyroiditis, pemphigus vulgaris, pernicious anemia, polychondritis,polymyositis and dermatomyositis, primary biliary cirrhosis,polyarteritis nodosa, polyglandular syndromes, polymyalgia rheumatica,primary agammaglobulinemia, Raynaud phenomenon, Reiter's syndrome,rheumatic fever, sarcoidosis, scleroderma, Sjogren's syndrome, stiffperson syndrome, Takayasu's arteritis, temporal arteritis, ulcerativecolitis, uveitis, vasculitis, vitiligo, vulvodynia, and Wegener'sgranulomatosis.

In some embodiments, the hematopoietic stem cells are autologous withrespect to the patient. In some embodiments, the hematopoietic stemcells are allogeneic with respect to the patient, and may be, forexample, HLA-matched with respect to the patient.

In some embodiments, the hematopoietic stem cells have been geneticallymodified to disrupt an endogenous gene, such as a gene encoding a majorhistocompatibility complex protein. The hematopoietic stem cells may begenetically modified to disrupt an endogenous by way of, for example, aCRISPR-associated protein, such as caspase 9, or another nucleasedescribed herein, such as a transcription activator-like effectornuclease, a meganuclease, or a zinc finger nuclease.

In some embodiments, the hematopoietic stem cells, or progeny thereof,maintain hematopoietic stem cell functional potential after two or moredays following infusion of the hematopoietic stem cells, or progenythereof, into the patient. In some embodiments, the hematopoietic stemcells, or progeny thereof, localize to hematopoietic tissue and/orreestablish hematopoiesis following infusion of the hematopoietic stemcells, or progeny thereof, into the patient. In some embodiments, uponinfusion into the patient, the hematopoietic stem cells, or progenythereof, give rise to recovery of a population of cells selected fromthe group consisting of megakaryocytes, thrombocytes, platelets,erythrocytes, mast cells, myeoblasts, basophils, neutrophils,eosinophils, microglia, granulocytes, monocytes, osteoclasts,antigen-presenting cells.

In another aspect, the disclosure relates to a method of mobilizingCD34^(dim) cells from the bone marrow of a human donor into peripheralblood, the method comprising administering to the donor (i) a CXCR2agonist selected from the group consisting of Gro-β, Gro-β T, andvariants thereof at a dose of from about 50 μg/kg to about 1,000 μg/kgand (ii) a CXCR4 antagonist.

In another aspect, the disclosure relates to a method of performing anallogeneic hematopoietic stem cell transplant in a patient in needthereof, the method comprising infusing into the patient atherapeutically effective amount of allogeneic hematopoietic stem cells,wherein the hematopoietic stem cells were mobilized from bone marrow ofa human donor into peripheral blood of the human donor by a methodcomprising administering to the donor (i) a CXCR2 agonist selected fromthe group consisting of Gro-β, Gro-β T, and variants thereof at a doseof from about 50 μg/kg to about 1,000 μg/kg and (ii) a CXCR4 antagonist.

In another aspect, the disclosure relates to a method of preventing,reducing the risk of developing, or reducing the severity of apost-transplant infection in a patient in need thereof, the methodcomprising infusing into the patient a therapeutically effective amountof hematopoietic stem cells, wherein the hematopoietic stem cells weremobilized from bone marrow of a human donor into peripheral blood of thehuman donor by a method comprising administering to the human donor (i)a CXCR2 agonist selected from the group consisting of Gro-β, Gro-β T,and variants thereof at a dose of from about 50 μg/kg to about 1,000μg/kg and (ii) a CXCR4 antagonist.

In another aspect, the disclosure relates to a method of preventing,reducing the risk of developing, or reducing the severity of graftversus host disease (GVHD) in a patient in need thereof, the methodcomprising infusing into the patient a therapeutically effective amountof hematopoietic stem cells, wherein the hematopoietic stem cells weremobilized from bone marrow of a human donor into peripheral blood of thehuman donor by a method comprising administering to the human donor (i)a CXCR2 agonist selected from the group consisting of Gro-β, Gro-β T,and variants thereof at a dose of from about 50 μg/kg to about 1,000μg/kg and (ii) a CXCR4 antagonist.

In certain embodiments, the CD34^(dim) cells are present in a higheramount in the peripheral blood than if the hematopoietic stem cells weremobilized using the CXCR4 antagonist alone. In certain embodiments, theCD34^(dim) cells are capable of suppressing alloreactive T lymphocyteproliferation when administered to a recipient.

In certain embodiments, the CXCR2 agonist is Gro-β T. In certainembodiments, the CXCR2 agonist is administered to the donor at a dose offrom about 100 μg/kg to about 250 μg/kg. In certain embodiments, theCXCR2 agonist is administered to the donor at a dose of from about 125μg/kg to about 225 μg/kg. In certain embodiments, the CXCR2 agonist isadministered to the donor at a dose of about 150 μg/kg. In certainembodiments, the CXCR2 agonist is administered intravenously to thedonor.

In certain embodiments, the CXCR4 antagonist is administeredsubcutaneously to the donor. In certain embodiments, the CXCR4antagonist is plerixafor or a pharmaceutically acceptable salt thereof.In certain embodiments, the plerixafor or pharmaceutically acceptablesalt thereof is administered to the donor at a dose of from about 50μg/kg to about 500 μg/kg. In certain embodiments, the plerixafor orpharmaceutically acceptable salt thereof is administered to the donor ata dose of from about 200 μg/kg to about 300 μg/kg. In certainembodiments, the plerixafor or pharmaceutically acceptable salt thereofis administered to the donor at a dose of about 240 μg/kg.

In certain embodiments, the method further includes testing a sample ofperipheral blood for the presence of CD34^(dim) cells and releasing thesample for ex vivo expansion of the CD34^(dim) cells.

In certain embodiments, the disclosure relates to the population ofCD34^(dim) cells derived from any of the above methods, or a compositioncomprising the same.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1A provides a graph showing that, when Gro-β T is co-administeredwith plerixafor (AMD3100) in mice, a synergistic increase inmobilization results, and grafts are enriched in highly engraftable,long-term hematopoietic stem cells (LT-HSC=Lin-c-kit⁺ Sca-1⁺ CD150⁺CD48⁺). FIG. 1B provides a graph showing that grafts containing cellsmobilized by Gro-β T and plerixaflor led to higher relative numbers ofcompetitive repopulating units (CRU) at week 16 than did graftscontaining cells mobilized by G-CSF alone.

FIG. 2A is a graph demonstrating the pharmacokinetic profile of variousdosages of Gro-β T when administered intravenously to Rhesus monkeys.FIG. 2B is a graph demonstrating the pharmacokinetic profile of variousdosages of Gro-β T when administered subcutaneously to Rhesus monkeys.In all experiments, Gro-β T was administered to subjects concurrentlywith plerixafor.

FIG. 3A shows a series of graphs demonstrating the mobilization responseof leukocytes (white blood cells, “WBCs”) to various dosages of Gro-β Tupon intravenous administration to Rhesus monkeys. Leukocyte response isshown both in terms of the quantity of cells mobilized (top) and thefold change in leukocyte density relative to baseline leukocyte densityprior to administration (bottom). FIG. 3B shows a series of graphsdemonstrating the mobilization response of leukocytes (white bloodcells, “WBCs”) to various dosages of Gro-β T upon subcutaneousadministration to Rhesus monkeys. Leukocyte response is shown both interms of the quantity of cells mobilized (top) and the fold change inleukocyte density relative to baseline leukocyte density prior toadministration (bottom). In all experiments, Gro-β T was administered tosubjects concurrently with plerixafor.

FIG. 4A shows a series of graphs demonstrating the mobilization responseof neutrophils to various dosages of Gro-β T upon intravenousadministration to Rhesus monkeys. Neutrophil response is shown both interms of the quantity of cells mobilized (top) and the fold change inneutrophil density relative to baseline neutrophil density prior toadministration (bottom). FIG. 4B shows a series of graphs demonstratingthe mobilization response of neutrophils to various dosages of Gro-β Tupon subcutaneous administration to Rhesus monkeys. Neutrophil responseis shown both in terms of the quantity of cells mobilized (top) and thefold change in neutrophil density relative to baseline neutrophildensity prior to administration (bottom). In all experiments, Gro-β Twas administered to subjects concurrently with plerixafor.

FIG. 5A shows a series of graphs demonstrating the mobilization responseof lymphocytes to various dosages of Gro-β T upon intravenousadministration to Rhesus monkeys. Lymphocyte response is shown both interms of the quantity of cells mobilized (top) and the fold change inlymphocyte density relative to baseline lymphocyte density prior toadministration (bottom). FIG. 5B shows a series of graphs demonstratingthe mobilization response of lymphocytes to various dosages of Gro-β Tupon subcutaneous administration to Rhesus monkeys. Lymphocyte responseis shown both in terms of the quantity of cells mobilized (top) and thefold change in lymphocyte density relative to baseline lymphocytedensity prior to administration (bottom). In all experiments, Gro-β Twas administered to subjects concurrently with plerixafor.

FIG. 6A shows a series of graphs demonstrating the mobilization responseof monocytes to various dosages of Gro-β T upon intravenousadministration to Rhesus monkeys. Monocyte response is shown both interms of the quantity of cells mobilized (top) and the fold change inmonocyte density relative to baseline monocyte density prior toadministration (bottom). FIG. 6B shows a series of graphs demonstratingthe mobilization response of monocytes to various dosages of Gro-β Tupon subcutaneous administration to Rhesus monkeys. Monocyte response isshown both in terms of the quantity of cells mobilized (top) and thefold change in monocyte density relative to baseline monocyte densityprior to administration (bottom). In all experiments, Gro-β T wasadministered to subjects concurrently with plerixafor.

FIG. 7A shows a series of graphs demonstrating the mobilization responseof CD34⁺ cells to various dosages of Gro-β T upon intravenousadministration to Rhesus monkeys. CD34⁺ cell response is shown both interms of the frequency of CD34⁺ cells in the sample obtained fromperipheral blood of the subjects (top) and the fold change in CD34⁺ cellfrequency relative to baseline CD34⁺ cell frequency prior toadministration (bottom). FIG. 7B shows a series of graphs demonstratingthe mobilization response of CD34⁺ cells to various dosages of Gro-β Tupon subcutaneous administration to Rhesus monkeys.CD34^(+ cell response is shown both in terms of the frequency of CD)34⁺cells in the sample obtained from peripheral blood of the subjects (top)and the fold change in CD34⁺ cell frequency relative to baseline CD34⁺cell frequency prior to administration (bottom). In all experiments,Gro-β T was administered to subjects concurrently with plerixafor.

FIG. 8A shows a series of graphs demonstrating the mobilization responseof CD34⁺ cells to various dosages of Gro-β T upon intravenousadministration to Rhesus monkeys. CD34⁺ cell response is shown both interms of the quantity of cells mobilized (top) and the fold change inCD34⁺ cell density relative to baseline CD34⁺ cell density prior toadministration (bottom). FIG. 8B shows a series of graphs demonstratingthe mobilization response of CD34⁺ cells to various dosages of Gro-β Tupon subcutaneous administration to Rhesus monkeys. CD34⁺ cell responseis shown both in terms of the quantity of cells mobilized (top) and thefold change in CD34⁺ cell density relative to baseline CD34⁺ celldensity prior to administration (bottom). In all experiments, Gro-β Twas administered to subjects concurrently with plerixafor.

FIG. 9A shows a series of graphs demonstrating the mobilization responseof hematopoietic stem cells (CD34⁺ CD90⁺ CD45RA⁻ cells) to variousdosages of Gro-β T upon intravenous administration to Rhesus monkeys.CD34⁺ CD90⁺ CD45RA⁻ cell response is shown both in terms of thefrequency of CD34⁺ CD90⁺ CD45RA⁻ cells in the sample obtained fromperipheral blood of the subjects (top) and the fold change in CD34⁺CD90⁺ CD45RA⁻ cell frequency relative to baseline CD34⁺ CD90⁺ CD45RA⁻cell frequency prior to administration (bottom). FIG. 9B shows a seriesof graphs demonstrating the mobilization response of hematopoietic stemcells (CD34⁺ CD90⁺ CD45RA⁻ cells) to various dosages of Gro-β T uponsubcutaneous administration to Rhesus monkeys. CD34⁺ CD90⁺ CD45RA⁻ cellresponse is shown both in terms of the frequency of CD34⁺ CD90⁺ CD45RA⁻cells in the sample obtained from peripheral blood of the subjects (top)and the fold change in CD34⁺ CD90⁺ CD45RA⁻ cell frequency relative tobaseline CD34⁺ CD90⁺ CD45RA⁻ cell frequency prior to administration(bottom). In all experiments, Gro-β T was administered to subjectsconcurrently with plerixafor.

FIG. 10A shows a series of graphs demonstrating the mobilizationresponse of hematopoietic stem cells (CD34⁺ CD90⁺ CD45RA⁻ cells) tovarious dosages of Gro-β T upon intravenous administration to Rhesusmonkeys. CD34⁺ CD90⁺ CD45RA⁻ cell response is shown both in terms of thequantity of cells mobilized (top) and the fold change in CD34⁺CD90⁺CD45RA⁻ cell density relative to baseline CD34⁺ CD90⁺ CD45RA⁻ celldensity prior to administration (bottom). FIG. 10B shows a series ofgraphs demonstrating the mobilization response of hematopoietic stemcells (CD34⁺ CD90⁺ CD45RA⁻ cells) to various dosages of Gro-β T uponsubcutaneous administration to Rhesus monkeys. CD34⁺ cell response isshown both in terms of the quantity of cells mobilized (top) and thefold change in CD34⁺ CD90⁺ CD45RA⁻ cell density relative to baselineCD34⁺ CD90⁺ CD45RA⁻ cell density prior to administration (bottom). Inall experiments, Gro-β T was administered to subjects concurrently withplerixafor.

FIG. 11 shows a series of graphs demonstrating the increase in thequantity of colony-forming units (CFU) of hematopoietic stem cellsachieved by the intravenous administration of various dosages of Gro-β Tto Rhesus monkeys. CFR response is shown both in terms of theconcentration of CFUs (top) and the fold change in CFU concentrationrelative to baseline CFU concentration prior to administration (bottom).In all experiments, Gro-β T was administered to subjects concurrentlywith plerixafor.

FIG. 12A shows a series of graphs demonstrating the response of plasmamatrix metalloproteinase 9 (MMP9) to various dosages of Gro-β T uponintravenous administration to Rhesus monkeys. Plasma MMP9 response isshown both in terms of absolute concentration (top) and the fold changein plasma MMP9 concentration relative to baseline MMP9 concentrationprior to administration (bottom). FIG. 12B shows a series of graphsdemonstrating the response of plasma MMP9 to various dosages of Gro-β Tupon subcutaneous administration to Rhesus monkeys. Plasma MMP9 responseis shown both in terms of absolute concentration (top) and the foldchange in plasma MMP9 concentration relative to baseline MMP9concentration prior to administration (bottom). In all experiments,Gro-β T was administered to subjects concurrently with plerixafor.

FIG. 13A shows a series of graphs demonstrating the response of plasmatissue inhibitor of matrix metalloproteinase 1 (TIMP-1) to variousdosages of Gro-β T upon intravenous administration to Rhesus monkeys.Plasma TIMP-1 response is shown both in terms of absolute concentration(top) and the fold change in plasma TIMP-1 concentration relative tobaseline TIMP-1 concentration prior to administration (bottom). FIG. 13Bshows a series of graphs demonstrating the response of plasma TIMP-1 tovarious dosages of Gro-β T upon subcutaneous administration to Rhesusmonkeys. Plasma TIMP-1 response is shown both in terms of absoluteconcentration (top) and the fold change in plasma TIMP-1 concentrationrelative to baseline TIMP-1 concentration prior to administration(bottom). In all experiments, Gro-β T was administered to subjectsconcurrently with plerixafor.

FIG. 14A is a graph showing the response of the molar ratio of plasmaMMP9 to plasma TIMP-1 to various dosages of Gro-β T upon intravenousadministration to Rhesus monkeys. FIG. 14B is a graph showing theresponse of the molar ratio of plasma MMP9 to plasma TIMP-1 to variousdosages of Gro-β T upon subcutaneous administration to Rhesus monkeys.In all experiments, Gro-β T was administered to subjects concurrentlywith plerixafor.

FIG. 15 provides representative flow plots from blood samples taken fourhours post-mobilization from Rhesus monkeys. Intravenous administrationof 450 μg/kg Gro-β T and subcutaneous administration of 1 mg/kgplerixafor (AMD3100) leads to the mobilization of a population ofCD34^(dim) cells.

FIG. 16 provides representative flow plots from blood samples taken atbaseline versus four hours post-mobilization from Rhesus monkeys.Mobilization was induced by (1) intravenous administration of 450 μg/kgGro-β T and subcutaneous administration of 1 mg/kg plerixafor (AMD3100)or (2) subcutaneous administration of 1 mg/kg plerixafor (AMD3100). Thecombination of Gro-β T and plerixafor (as compared to plerixafor alone)leads to enhanced mobilization of CD34^(dim) cells.

FIG. 17 is a graph quantifying the concentration of CD34^(dim) cells inperipheral blood from untreated Rhesus monkeys (“Unmobilized”), Rhesusmonkeys that have been treated with intravenous administration of 450μg/kg Gro-β T and subcutaneous administration of 1 mg/kg plerixafor(“Gro-β T+plerixafor”), Rhesus monkeys that have been treated withsubcutaneous administration of 1 mg/kg plerixafor (“plerixafor”) andRhesus monkeys that have been treated with subcutaneous administrationof 50 μg/kg (q.d.×5) G-CSF (“G-CSF”). CD34^(dim) cells were present at asignificantly higher concentration in blood mobilized using Gro-β T plusplerixafor.

FIG. 18 is a graph depicting the composition of unmobilized cells andgrafts mobilized by G-CSF, Gro-β T and AMD3100 and AMD3100 alone. Asshown, grafts mobilized using Gro-β T and AMD3100 show a 3 fold increasein CD34^(dim) cells and a 3 fold increase in T-cells as compared tografts mobilized using G-CSF.

FIG. 19 provides graphs showing that Gro-β T and AMD3100 mobilizedCD34^(dim) cells suppressed T-cell proliferation as measured bycarboxyfluorescein succinimidyl ester (CFSE) staining after four days inculture. “Beads” indicates stimulation of T-cells usinganti-CD2/CD3/CD28 coated microbeads.

FIG. 20 provides a survival curve showing that by day 24, all mice(13/13) transplanted with unmobilized PBMCs had died of aGVHD comparedto 5/16 mice transplanted with AMD3100 mobilized peripheral blood, 3/16mice transplanted with G-CSF mobilized PBMCs and none of the micetransplanted with Gro-β T and AMD3100 mobilized PBMCs. At day 60post-transplant, 15/16 mice transplanted with Gro-β T and AMD3100remained alive, whereas only 10/16 mice transplanted with AMD3100mobilized PBMCs and 11/16 mice transplanted with G-CSF mobilized PBMCsremained alive. p<0.0001 (comparing Gro-β T and AMD3100 v. unmobilized)and p<0.05 (comparing Gro-β T and AMD3100 v. AMD3100 alone).

FIG. 21A shows rhesus CD45+CD3+ T-cell numbers in mice at day 14post-transplant with unmobilized PBMCs, PBMCs mobilized with Gro-β T andplerixafor (i.e., AMD3100), PBMCs mobilized with plerixafor alone, andPBMCs mobilized with G-CSF alone. FIG. 21B shows T-cell numbers in miceat day 14 post-transplant with unmobilized PBMCs, Gro-β T and plerixafor(i.e., AMD3100) mobilized PBMCs, and Gro-β T and plerixafor mobilizedPBMCs that have been depleted of CD34dim cells. FIG. 21C provides asurvival curve of mice transplanted with unmobilized PBMCs, Gro-β T andAMD3100 mobilized PBMCs, and Gro-β T and AMD3100 mobilized PBMCs thathave been depleted of CD34dim cells.

DETAILED DESCRIPTION

The present invention provides compositions and methods for mobilizinghematopoietic stem and progenitor cells in a subject. For example, thesubject may be a hematopoietic stem and progenitor cell donor (i.e., adonor), such as a mammalian donor (e.g., a human donor). Thecompositions and methods described herein can additionally be used forthe treatment of one or more stem cell disorders in a patient, such as ahuman patient. Using the compositions and methods described herein, aC-X-C chemokine receptor type 2 (CXCR2) agonist, such as Gro-β or avariant thereof, such as a truncated form of Gro-β (e.g., Gro-β T), asdescribed herein, optionally in combination with a C-X-C chemokinereceptor type 4 (CXCR4) antagonist, such as1,1′-[1,4-phenylenebis(methylene)]-bis-1,4,8,11-tetra-azacyclotetradecaneor a variant thereof, may be administered to a donor, as describedherein, in amounts sufficient to mobilize hematopoietic stem andprogenitor cells. The compositions and methods described herein arecapable of mobilizing hematopoietic stem and progenitor cells from astem cell niche within a donor into circulating peripheral blood whilereducing the mobilization of other cells of the hematopoietic lineage,such as leukocytes, neutrophils, lymphocytes, and monocytes. Thecompositions and methods described herein thus enable the selectivemobilization of hematopoietic stem and progenitor cells in a donor,which may then be isolated from a donor for therapeutic use.

The invention is based, in part, on the discovery that administration ofa CXCR2 agonist, such as Gro-β, Gro-β T, or a variant thereof,optionally in combination with a CXCR4 antagonist, such as plerixafor ora pharmaceutically acceptable salt thereof, at particular doses canprovide the important clinical benefit of mobilizing populations ofcells that are enriched in hematopoietic stem cells relative to othercell types, such as leukocytes, neutrophils, and monocytes. This abilityis advantageous, as these other cell types may be undesirable foradministration to a human patient undergoing hematopoietic stem celltransplant therapy. Thus, the populations of mobilized hematopoieticstem and progenitor cells produced using the compositions and methodsdescribed herein are particularly suitable for hematopoietic stem celltransplantation therapy.

Following mobilization, the hematopoietic stem or progenitor cells maybe isolated for ex vivo expansion and/or for therapeutic use. In someembodiments, upon collection of the mobilized hematopoietic stem and/orprogenitor cells, the withdrawn cells may be infused into a patient,such as the donor or another subject (e.g., a subject that isHLA-matched to the donor) for the treatment of one or more pathologiesof the hematopoietic system. Additionally or alternatively, themobilized cells may be withdrawn and then expanded ex vivo, such as bycontacting the cells with an aryl hydrocarbon receptor antagonist, so asto produce a population of hematopoietic stem cells having a sufficientquantity of cells for transplantation.

As described herein, hematopoietic stem cells are capable ofdifferentiating into a multitude of cell types in the hematopoieticlineage, and can thus be administered to a patient in order to populateor repopulate a cell type that is defective or deficient in the patient.The patient may be one, for example, that is suffering from one or moreblood disorders, such as an autoimmune disease, cancer,hemoglobinopathy, or other hematopoietic pathology, and is therefore inneed of hematopoietic stem cell transplantation. The invention thusprovides methods of treating a variety of hematopoietic conditions, suchas sickle cell anemia, thalassemia, Fanconi anemia, Wiskott-Aldrichsyndrome, adenosine deaminase deficiency-severe combinedimmunodeficiency, metachromatic leukodystrophy, Diamond-Blackfan anemiaand Schwachman-Diamond syndrome, human immunodeficiency virus infection,and acquired immune deficiency syndrome, as well as cancers andautoimmune diseases, among others.

The sections that follow provide a description of CXCR4 antagonists andCXCR2 agonists that can be administered to a donor so as to inducemobilization of a population of hematopoietic stem or progenitor cellsfrom a stem cell niche into peripheral blood, from which the cells maysubsequently be isolated and infused into a patient for the treatment,for example, of one or more stem cell disorders, such as a cancer,autoimmune disease, of metabolic disorder described herein. Thefollowing sections additionally describe methods of determining whetherpopulations of cells mobilized with a CXCR2 agonist and/or a CXCRantagonist are suitable for release for ex vivo expansion and/or fortherapeutic applications.

Definitions

As used herein, the term “about” refers to a value that is within 10%above or below the value being described. For example, the term “about 5nM” indicates a range of from 4.5 nM to 5.5 nM.

As used herein, the terms “acquire” and “acquiring” means obtainingpossession of a physical entity, or a value, such as a numerical value,directly acquiring or indirectly acquiring the physical entity or value.“Directly acquiring” means performing a process (e.g., performing anassay or test on a sample or analyzing a sample) to obtain the physicalentity or value. “Indirectly acquiring” refers to receiving the physicalentity or value from another party or source (e.g., a third partylaboratory that directly acquired the physical entity or value).Directly acquiring a physical entity includes performing a process,e.g., analyzing a sample, such as a sample of hematopoietic cellsisolated from a donor that has undergone or is undergoing ahematopoietic stem cell mobilization regimen described herein. Directlyacquiring a value includes performing a process, such as an assay, on asample or another substance, e.g., performing an analytical processwhich includes determining the quantity of hematopoietic stem cells in asample, the ratio of hematopoietic stem cells to cells of another typewithin the hematopoietic lineage, or the frequency of hematopoietic stemcells among the total quantity of cells in a sample.

As used herein, the term “affinity” refers to the strength of thenon-covalent interaction between two or more molecules, such as two ormore proteins (e.g., a metalloproteinase and an endogenous inhibitorthereof as described herein). Affinity can be expressed quantitatively,for example, as an equilibrium dissociation constant (K_(d)) or, incases in which one of the binding partners is an enzyme, as aninhibition constant (K_(i)). Binding affinity can be determined usingstandard techniques, such as enzyme-linked immunosorbent assays (ELISA),surface plasmon resonance assays, and isothermal titration calorimetryassays, among others.

As used herein, the term “antibody” refers to an immunoglobulin moleculethat specifically binds to, or is immunologically reactive with, aparticular antigen, and includes polyclonal, monoclonal, geneticallyengineered, and otherwise modified forms of antibodies, including butnot limited to chimeric antibodies, humanized antibodies,heteroconjugate antibodies (e.g., bi- tri- and quad-specific antibodies,diabodies, triabodies, and tetrabodies), and antigen binding fragmentsof antibodies, including, for example, Fab′, F(ab′)₂, Fab, Fv, rlgG, andscFv fragments. Unless otherwise indicated, the term “monoclonalantibody” (mAb) is meant to include both intact molecules, as well asantibody fragments (including, for example, Fab and F(ab′)₂ fragments)that are capable of specifically binding to a target protein. As usedherein, the Fab and F(ab′)₂ fragments refer to antibody fragments thatlack the Fc fragment of an intact antibody. Examples of these antibodyfragments are described herein.

The term “antigen-binding fragment,” as used herein, refers to one ormore fragments of an antibody that retain the ability to specificallybind to a target antigen. The antigen-binding function of an antibodycan be performed by fragments of a full-length antibody.

The antibody fragments can be, for example, a Fab, F(ab′)₂, scFv,diabody, a triabody, an affibody, a nanobody, an aptamer, or a domainantibody. Examples of binding fragments encompassed of the term“antigen-binding fragment” of an antibody include, but are not limitedto: (i) a Fab fragment, a monovalent fragment consisting of the V_(L),V_(H), C_(L), and C_(H)1 domains; (ii) a F(ab′)₂ fragment, a bivalentfragment containing two Fab fragments linked by a disulfide bridge atthe hinge region; (iii) a Fd fragment consisting of the V_(H) and C_(H)1domains; (iv) a Fv fragment consisting of the V_(L) and V_(H) domains ofa single arm of an antibody, (v) a dAb including V_(H) and V_(L)domains; (vi) a dAb fragment that consists of a V_(H) domain (see, e.g.,Ward et al. (1989) Nature 341:544-546); (vii) a dAb which consists of aV_(H) or a V_(L) domain; (viii) an isolated complementarity determiningregion (CDR); and (ix) a combination of two or more (e.g., two, three,four, five, or six) isolated CDRs which may optionally be joined by asynthetic linker. Furthermore, although the two domains of the Fvfragment, V_(L) and V_(H), are coded for by separate genes, they can bejoined, using recombinant methods, by a linker that enables them to bemade as a single protein chain in which the V_(L) and V_(H) regions pairto form monovalent molecules (known as single chain Fv (scFv); see, forexample, Bird et al. (1988) Science 242:423-426 and Huston et al. (1988)Proc. Natl. Acad. Sci. USA 85:5879-5883). These antibody fragments canbe obtained using conventional techniques known to those of skill in theart, and the fragments can be screened for utility in the same manner asintact antibodies. Antigen-binding fragments can be produced byrecombinant DNA techniques, enzymatic or chemical cleavage of intactimmunoglobulins, or, in certain cases, by chemical peptide synthesisprocedures known in the art.

As used herein, the term “bispecific antibody” refers to, for example, amonoclonal, often a human or humanized antibody that is capable ofbinding at least two different antigens.

As used herein the term “CD34^(dim) cells” refers to a population ofcells, of which at least a portion of the population expresses themarkers CD34, CD11b, and CD45 and does not substantially express themarkers CD3, CD8, or CD20, wherein CD34 and CD45 are expressed at arelatively low level. This cell population exhibits characteristics ofmonocytes, for example, the ability to suppress alloreactive Tlymphocyte proliferation. (D'Aveni et al. (2015) Science TranslationalMedicine 7(281):1-12. In some embodiments, a population of CD34^(dim)cells may be CD14+.

The person of skill can readily recognize a CD34^(dim) cell populationwhen viewing a flow cytometry plot as a population of cells that isCD34⁺ but has a brightness between the base level of fluorescence (e.g.,autofluorescence) in the cell population being examined and thebrightness of a CD34⁺ bright cell (e.g., a hematopoietic stem celland/or a CD34⁺ CD90⁺ cell) population. For example, in certainembodiments, a CD34^(dim) cell population exhibits between 5% and 95% ofthe brightness of a CD34⁺ bright cell (e.g., a hematopoietic stem cell)population, but is brighter than a CD34⁻ cell population.

In certain embodiments, CD34^(dim) cells exhibit between 10% and 90%,10% and 80%, 10% and 70%, 10% and 60%, 10% and 50%, 10% and 40%, 10% and30%, 10% and 20%, 20% and 90%, 20% and 80%, 20% and 70%, 20% and 60%,20% and 50%, 20% and 40%, 20% and 30%, 30% and 90%, 30% and 80%, 30% and70%, 30% and 60%, 30% and 50%, 30% and 40%, 40% and 90%, 40% and 80%,40% and 70%, 40% and 60%, 40% and 50%, 50% and 90%, 50% and 80%, 50% and70%, 50% and 60%, 60% and 90%, 60% and 80%, 60% and 70%, 70% and 90%,70% and 80%, or 80% and 90%, of the brightness of CD34⁺ bright cells(e.g., hematopoietic stem cells), but is brighter than CD34⁻ cells. Incertain embodiments, the CD34^(dim) cells are at least 5% brighter, atleast 10% brighter, at least 20% brighter or at least 30% brighter thanCD34⁻ cells, but is less bright than CD34⁺ bright cells (e.g., ahematopoietic stem cells).

In certain embodiments, CD34^(dim) cells are identified in a cell sampleby using flow cytometry with magnetic beads instead of fluorescence. Themagnetic beads will selectively pull down CD34⁺ bright cells, leavingCD34^(dim) cells in the cell sample.

In certain embodiments, CD34^(dim) cells are identified by measuring thenumber of copies of CD34 expressed by the cells. For example, CD34^(dim)cells can exhibit between 5% and 95% of the number of copies of CD34 ascompared to CD34⁺ bright cells (e.g., hematopoietic stem cells), orbetween 10% and 90%, 10% and 80%, 10% and 70%, 10% and 60%, 10% and 50%,10% and 40%, 10% and 30%, 10% and 20%, 20% and 90%, 20% and 80%, 20% and70%, 20% and 60%, 20% and 50%, 20% and 40%, 20% and 30%, 30% and 90%,30% and 80%, 30% and 70%, 30% and 60%, 30% and 50%, 30% and 40%, 40% and90%, 40% and 80%, 40% and 70%, 40% and 60%, 40% and 50%, 50% and 90%,50% and 80%, 50% and 70%, 50% and 60%, 60% and 90%, 60% and 80%, 60% and70%, 70% and 90%, 70% and 80%, or 80% and 90% of the number of copies ofCD34 as compared to CD34⁺ bright cells (e.g., hematopoietic stem cells).

As used herein, the term “complementarity determining region” (CDR)refers to a hypervariable region found both in the light chain and theheavy chain variable domains of an antibody. The more highly conservedportions of variable domains are referred to as framework regions (FRs).The amino acid positions that delineate a hypervariable region of anantibody can vary, depending on the context and the various definitionsknown in the art. Some positions within a variable domain may be viewedas hybrid hypervariable positions in that these positions can be deemedto be within a hypervariable region under one set of criteria whilebeing deemed to be outside a hypervariable region under a different setof criteria. One or more of these positions can also be found inextended hypervariable regions. The antibodies described herein maycontain modifications in these hybrid hypervariable positions. Thevariable domains of native heavy and light chains each contain fourframework regions that primarily adopt a n-sheet configuration,connected by three CDRs, which form loops that connect, and in somecases form part of, the β-sheet structure. The CDRs in each chain areheld together in close proximity by the framework regions in the orderFR1-CDR1-FR2-CDR2-FR3-CDR3-FR4 and, with the CDRs from the otherantibody chains, contribute to the formation of the target binding siteof antibodies (see Kabat et al., Sequences of Proteins of ImmunologicalInterest, National Institute of Health, Bethesda, Md., 1987). As usedherein, numbering of immunoglobulin amino acid residues is performedaccording to the immunoglobulin amino acid residue numbering system ofKabat et al., unless otherwise indicated.

As used herein in the context of the administration of one or moreagents to a subject, the term “completion of administration” refers tothe point in time by which the one or more agents have been administeredto the subject in their entirety. In some embodiments, an agent asdescribed herein, such as a CXCR4 antagonist (e.g., plerixafor or avariant thereof) and/or a CXCR2 agonist (e.g., Gro-β or a variant ortruncation thereof, such as Gro-β T) can be administered to a subjectover a period of time, for example, by intravenous or subcutaneousinjection. An agent is considered to have “completed administration”once the prescribed dosage of the agent has been administered to thesubject in its entirety. In the case of the administration of multipleagents to a subject, such as both a CXCR4 antagonist (e.g., plerixaforor a variant thereof) and a CXCR2 agonist (e.g., Gro-β or a variant ortruncation thereof, such as Gro-β T), the agents are considered to have“completed administration” once the prescribed dosages of all agents ina particular regimen have been administered to the subject in theirentirety.

As used herein, the terms “conservative mutation,” “conservativesubstitution,” or “conservative amino acid substitution” refer to asubstitution of one or more amino acids for one or more different aminoacids that exhibit similar physicochemical properties, such as polarity,electrostatic charge, and steric volume. These properties are summarizedfor each of the twenty naturally-occurring amino acids in TABLE 1 below.

TABLE 1 Representative physicochemical properties of naturally-occurringamino acids Electrostatic character at 3 Letter 1 Letter Side-chainphysiological Steric Amino Acid Code Code Polarity pH (7.4) Volume^(†)Alanine Ala A nonpolar neutral small Arginine Arg R polar cationic largeAsparagine Asn N polar neutral intermediate Aspartic acid Asp D polaranionic intermediate Cysteine Cys C nonpolar neutral intermediateGlutamic acid Glu E polar anionic intermediate Glutamine Gln Q polarneutral intermediate Glycine Gly G nonpolar neutral small Histidine HisH polar Both neutral large and cationic forms in equilibrium at pH 7.4Isoleucine Ile I nonpolar neutral large Leucine Leu L nonpolar neutrallarge Lysine Lys K polar cationic large Methionine Met M nonpolarneutral large Phenylalanine Phe F nonpolar neutral large Proline Pro Pnon-polar neutral intermediate Serine Ser S polar neutral smallThreonine Thr T polar neutral intermediate Tryptophan Trp W nonpolarneutral bulky Tyrosine Tyr Y polar neutral large Valine Val V nonpolarneutral intermediate ^(†)based on volume in A3: 50-100 is small, 100-500is intermediate, 150-200 is large and >200 is bulky

From this table it is appreciated that the conservative amino acidfamilies include, e.g., (i) G, A, V, L, I, P, and M; (ii) D and E; (iii)C, S and T; (iv) H, K and R; (v) N and Q; and (vi) F, Y and W. Aconservative mutation or substitution is therefore one that substitutesone amino acid for a member of the same amino acid family (e.g., asubstitution of Ser for Thr or Lys for Arg).

As used herein, “CRU (competitive repopulating unit)” refers to a unitof measure of long-term engrafting stem cells, which can be detectedafter in-vivo transplantation.

As used herein, the term “donor” refers to a subject, such as amammalian subject (e.g., a human subject) from which one or more cellsare isolated prior to administration of the cells, or progeny thereof,into a recipient. The one or more cells may be, for example, apopulation of hematopoietic stem or progenitor cells.

As used herein, the term “diabody” refers to a bivalent antibodycontaining two polypeptide chains, in which each polypeptide chainincludes V_(H) and V_(L) domains joined by a linker that is too short(e.g., a linker composed of five amino acids) to allow forintramolecular association of V_(H) and V_(L) domains on the samepeptide chain. This configuration forces each domain to pair with acomplementary domain on another polypeptide chain so as to form ahomodimeric structure. Accordingly, the term “triabody” refers totrivalent antibodies containing three peptide chains, each of whichcontains one V_(H) domain and one V_(L) domain joined by a linker thatis exceedingly short (e.g., a linker composed of 1-2 amino acids) topermit intramolecular association of V_(H) and V_(L) domains within thesame peptide chain. In order to fold into their native structures,peptides configured in this way typically trimerize so as to positionthe V_(H) and V_(L) domains of neighboring peptide chains spatiallyproximal to one another (see, for example, Holliger et al. (1993) Proc.Natl. Acad. Sci. USA 90:6444-48).

As used herein, the term “disrupt” with respect to a gene refers topreventing the formation of a functional gene product. A gene product isfunctional only if it fulfills its normal (wild-type) functions.Disruption of the gene prevents expression of a functional factorencoded by the gene and comprises an insertion, deletion, orsubstitution of one or more bases in a sequence encoded by the geneand/or a promoter and/or an operator that is necessary for expression ofthe gene in the animal. The disrupted gene may be disrupted by, e.g.,removal of at least a portion of the gene from a genome of the animal,alteration of the gene to prevent expression of a functional factorencoded by the gene, an interfering RNA, or expression of a dominantnegative factor by an exogenous gene. Materials and methods ofgenetically modifying hematopoietic stem/progenitor cells are detailedin U.S. Pat. No. 8,518,701; US 2010/0251395; and US 2012/0222143, thedisclosures of each of which are incorporated herein by reference intheir entirety (in case of conflict, the instant specification iscontrolling).

Various techniques known in the art can be used to inactivate genes tomake knockout animals and/or to introduce nucleic acid constructs intoanimals to produce founder animals and to make animal lines, in whichthe knockout or nucleic acid construct is integrated into the genome.Such techniques include, without limitation, pronuclear microinjection(U.S. Pat. No. 4,873,191), retrovirus mediated gene transfer into germlines (Van der Putten et al. (1985) Proc. Natl. Acad. Sci. USA,82:6148-6152), gene targeting into embryonic stem cells (Thompson et al.(1989) Cell, 56:313-321), electroporation of embryos (Lo (1983) Mol.Cell. Biol., 3:1803-1814), sperm-mediated gene transfer (Lavitrano etal. (2002) Proc. Natl. Acad. Sci. USA, 99:14230-14235; Lavitrano et al.(2006) Reprod. Fert. Develop., 18:19-23), and in vitro transformation ofsomatic cells, such as cumulus or mammary cells, or adult, fetal, orembryonic stem cells, followed by nuclear transplantation (Wilmut et al.(1997) Nature, 385:810-813; and Wakayama et al. (1998) Nature,394:369-374). Pronuclear microinjection, sperm mediated gene transfer,and somatic cell nuclear transfer are particularly useful techniques. Ananimal that is genomically modified is an animal wherein all of itscells have the genetic modification, including its germ line cells. Whenmethods are used that produce an animal that is mosaic in its geneticmodification, the animals may be inbred and progeny that are genomicallymodified may be selected. Cloning, for example, may be used to make amosaic animal if its cells are modified at the blastocyst state, orgenomic modification can take place when a single-cell is modified.Animals that are modified so they do not sexually mature can behomozygous or heterozygous for the modification, depending on thespecific approach that is used. If a particular gene is inactivated by aknock out modification, homozygosity would normally be required. If aparticular gene is inactivated by an RNA interference or dominantnegative strategy, then heterozygosity is often adequate.

As used herein, a “dual variable domain immunoglobulin” (“DVD-Ig”)refers to an antibody that combines the target-binding variable domainsof two monoclonal antibodies via linkers to create a tetravalent,dual-targeting single agent (see, for example, Gu et al. (2012) Meth.Enzymol., 502:25-41).

As used herein, the term “endogenous” describes a substance, such as amolecule, cell, tissue, or organ (e.g., a hematopoietic stem cell or acell of hematopoietic lineage, such as a megakaryocyte, thrombocyte,platelet, erythrocyte, mast cell, myeoblast, basophil, neutrophil,eosinophil, microglial cell, granulocyte, monocyte, osteoclast,antigen-presenting cell, macrophage, dendritic cell, natural killercell, T-lymphocyte, or B-lymphocyte) that is found naturally in aparticular organism, such as a human patient.

As used herein, the term “engraftment potential” is used to refer to theability of hematopoietic stem and progenitor cells to repopulate atissue, whether such cells are naturally circulating or are provided bytransplantation. The term encompasses all events surrounding or leadingup to engraftment, such as tissue homing of cells and colonization ofcells within the tissue of interest. The engraftment efficiency or rateof engraftment can be evaluated or quantified using any clinicallyacceptable parameter as known to those of skill in the art and caninclude, for example, assessment of competitive repopulating units(CRU); incorporation or expression of a marker in tissue(s) into whichstem cells have homed, colonized, or become engrafted; or by evaluationof the progress of a subject through disease progression, survival ofhematopoietic stem and progenitor cells, or survival of a recipient.Engraftment can also be determined by measuring white blood cell countsin peripheral blood during a post-transplant period. Engraftment canalso be assessed by measuring recovery of marrow cells by donor cells ina bone marrow aspirate sample.

As used herein, the term “exogenous” describes a substance, such as amolecule, cell, tissue, or organ (e.g., a hematopoietic stem cell or acell of hematopoietic lineage, such as a megakaryocyte, thrombocyte,platelet, erythrocyte, mast cell, myeoblast, basophil, neutrophil,eosinophil, microglial cell, granulocyte, monocyte, osteoclast,antigen-presenting cell, macrophage, dendritic cell, natural killercell, T-lymphocyte, or B-lymphocyte) that is not found naturally in aparticular organism, such as a human patient. Exogenous substancesinclude those that are provided from an external source to an organismor to cultured matter extracted therefrom.

As used herein, the term “framework region” or “FW region” includesamino acid residues that are adjacent to the CDRs of an antibody orantigen-binding fragment thereof. FW region residues may be present in,for example, human antibodies, humanized antibodies, monoclonalantibodies, antibody fragments, Fab fragments, single chain antibodyfragments, scFv fragments, antibody domains, and bispecific antibodies,among others.

As used herein, the term “hematopoietic progenitor cells” includespluripotent cells capable of differentiating into several cell types ofthe hematopoietic system, including, without limitation, granulocytes,monocytes, erythrocytes, megakaryocytes, B-cells and T-cells, amongothers. Hematopoietic progenitor cells are committed to thehematopoietic cell lineage and generally do not self-renew.Hematopoietic progenitor cells can be identified, for example, byexpression patterns of cell surface antigens, and include cells havingthe following immunophenotype: Lin⁻ KLS⁺ Flk2⁻ CD34⁺. Hematopoieticprogenitor cells include short-term hematopoietic stem cells,multi-potent progenitor cells, common myeloid progenitor cells,granulocyte-monocyte progenitor cells, and megakaryocyte-erythrocyteprogenitor cells. The presence of hematopoietic progenitor cells can bedetermined functionally, for example, by detecting colony-forming unitcells, e.g., in complete methylcellulose assays, or phenotypicallythrough the detection of cell surface markers using flow cytometry andcell sorting assays described herein and known in the art.

As used herein, the term “hematopoietic stem cells” (“HSCs”) refers toimmature blood cells having the capacity to self-renew and todifferentiate into mature blood cells containing diverse lineagesincluding but not limited to granulocytes (e.g., promyelocytes,neutrophils, eosinophils, basophils), erythrocytes (e.g., reticulocytes,erythrocytes), thrombocytes (e.g., megakaryoblasts, platelet producingmegakaryocytes, platelets), monocytes (e.g., monocytes, macrophages),dendritic cells, microglia, osteoclasts, and lymphocytes (e.g., NKcells, B-cells and T-cells). Such cells may include CD34⁺ cells. CD34⁺cells are immature cells that express the CD34 cell surface marker. Inhumans, CD34⁺ cells are believed to include a subpopulation of cellswith the stem cell properties defined above, whereas in mice, HSCs areCD34−. In addition, HSCs also refer to long term repopulating HSCs(LT-HSC) and short term repopulating HSCs (ST-HSC). LT-HSCs and ST-HSCsare differentiated, based on functional potential and on cell surfacemarker expression. For example, human HSCs are CD34⁺, CD38⁻, CD45RA⁻,CD90⁺, CD49F⁺, and lin⁻ (negative for mature lineage markers includingCD2, CD3, CD4, CD7, CD8, CD10, CD11B, CD19, CD20, CD56, CD235A). Inmice, bone marrow LT-HSCs are CD34-, SCA-1+, C-kit+, CD135−,Slamf1/CD150+, CD48−, and lin− (negative for mature lineage markersincluding Ter119, CD11b, Gr1, CD3, CD4, CD8, B220, IL7ra), whereasST-HSCs are CD34⁺, SCA-1⁺, C-kit⁺, CD135⁻, Slamf1/CD150⁺, and lin⁻(negative for mature lineage markers including Ter119, CD11b, Gr1, CD3,CD4, CD8, B220, IL7ra). In addition, ST-HSCs are less quiescent and moreproliferative than LT-HSCs under homeostatic conditions. However, LT-HSChave greater self-renewal potential (i.e., they survive throughoutadulthood, and can be serially transplanted through successiverecipients), whereas ST-HSCs have limited self-renewal (i.e., theysurvive for only a limited period of time, and do not possess serialtransplantation potential). Any of these HSCs can be used in the methodsdescribed herein. ST-HSCs are particularly useful because they arehighly proliferative and thus, can more quickly give rise todifferentiated progeny.

As used herein, the term “hematopoietic stem cell functional potential”refers to the functional properties of hematopoietic stem cells whichinclude 1) multi-potency (which refers to the ability to differentiateinto multiple different blood lineages including, but not limited to,granulocytes (e.g., promyelocytes, neutrophils, eosinophils, basophils),erythrocytes (e.g., reticulocytes, erythrocytes), thrombocytes (e.g.,megakaryoblasts, platelet producing megakaryocytes, platelets),monocytes (e.g., monocytes, macrophages), dendritic cells, microglia,osteoclasts, and lymphocytes (e.g., NK cells, B-cells and T-cells), 2)self-renewal (which refers to the ability of hematopoietic stem cells togive rise to daughter cells that have equivalent potential as the mothercell, and further that this ability can repeatedly occur throughout thelifetime of an individual without exhaustion), and 3) the ability ofhematopoietic stem cells or progeny thereof to be reintroduced into atransplant recipient whereupon they home to the hematopoietic stem cellniche and re-establish productive and sustained hematopoiesis.

As used herein, the terms “Major histocompatibility complex antigens”(“MHC”, also referred to as “human leukocyte antigens” (“HLA”) in thecontext of humans) refer to proteins expressed on the cell surface thatconfer a unique antigenic identity to a cell. MHC/HLA antigens aretarget molecules that are recognized by T cells and NK cells as beingderived from the same source of hematopoietic stem cells as the immuneeffector cells (“self”) or as being derived from another source ofhematopoietic reconstituting cells (“non-self”). Two main classes of HLAantigens are recognized: HLA class I and HLA class II. HLA class Iantigens (A, B, and C in humans) render each cell recognizable as“self,” whereas HLA class II antigens (DR, DP, and DQ in humans) areinvolved in reactions between lymphocytes and antigen presenting cells.Both have been implicated in the rejection of transplanted organs. Animportant aspect of the HLA gene system is its polymorphism. Each gene,MHC class I (A, B and C) and MEC class II (DP, DQ and DR) exists indifferent alleles. For example, two unrelated individuals may carryclass I HLA-B, genes B5, and Bw41, respectively. Allelic gene productsdiffer in one or more amino acids in the α and/or β domain(s). Largepanels of specific antibodies or nucleic acid reagents are used to typeHLA haplotypes of individuals, using leukocytes that express class I andclass II molecules. The genes commonly used for HLA typing are the sixMHC Class I and Class II proteins, two alleles for each of HLA-A; HLA-Band HLA-DR. The HLA genes are clustered in a “super-locus” present onchromosome position 6p21, which encodes the six classicaltransplantation HLA genes and at least 132 protein coding genes thathave important roles in the regulation of the immune system as well assome other fundamental molecular and cellular processes. The completelocus measures roughly 3.6 Mb, with at least 224 gene loci. One effectof this clustering is that “haplotypes”, i.e. the set of alleles presenton a single chromosome, which is inherited from one parent, tend to beinherited as a group. The set of alleles inherited from each parentforms a haplotype, in which some alleles tend to be associated together.Identifying a patient's haplotypes can help predict the probability offinding matching donors and assist in developing a search strategy,because some alleles and haplotypes are more common than others and theyare distributed at different frequencies in different racial and ethnicgroups.

As used herein, the term “HLA-matched” refers to a donor-recipient pairin which none of the HLA antigens are mismatched between the donor andrecipient, such as a donor providing a hematopoietic stem cell graft toa recipient in need of hematopoietic stem cell transplant therapy.HLA-matched (i.e., where all of the 6 alleles are matched)donor-recipient pairs have a decreased risk of graft rejection, asendogenous T cells and NK cells are less likely to recognize theincoming graft as foreign, and are thus less likely to mount an immuneresponse against the transplant.

As used herein, the term “HLA-mismatched” refers to a donor-recipientpair in which at least one HLA antigen, in particular with respect toHLA-A, HLA-B and HLA-DR, is mismatched between the donor and recipient,such as a donor providing a hematopoietic stem cell graft to a recipientin need of hematopoietic stem cell transplant therapy. In someembodiments, one haplotype is matched and the other is mismatched.HLA-mismatched donor-recipient pairs may have an increased risk of graftrejection relative to HLA-matched donor-recipient pairs, as endogenous Tcells and NK cells are more likely to recognize the incoming graft asforeign in the case of an HLA-mismatched donor-recipient pair, and suchT cells and NK cells are thus more likely to mount an immune responseagainst the transplant.

As used herein, the term “human antibody” refers to an antibody in whichsubstantially every part of the protein (for example, all CDRs,framework regions, C_(L), C_(H) domains (e.g., C_(H)1, C_(H)2, C_(H)3),hinge, and V_(L) and V_(H) domains) is substantially non-immunogenic inhumans, with only minor sequence changes or variations. A human antibodycan be produced in a human cell (for example, by recombinant expression)or by a non-human animal or a prokaryotic or eukaryotic cell that iscapable of expressing functionally rearranged human immunoglobulin (suchas heavy chain and/or light chain) genes. When a human antibody is asingle chain antibody, it can include a linker peptide that is not foundin native human antibodies. For example, an Fv can contain a linkerpeptide, such as two to about eight glycine or other amino acidresidues, which connects the variable region of the heavy chain and thevariable region of the light chain. Such linker peptides are consideredto be of human origin. Human antibodies can be made by a variety ofmethods known in the art including phage display methods using antibodylibraries derived from human immunoglobulin sequences. Human antibodiescan also be produced using transgenic mice that are incapable ofexpressing functional endogenous immunoglobulins, but which can expresshuman immunoglobulin genes (see, for example, PCT Publication Nos. WO1998/24893; WO 1992/01047; WO 1996/34096; WO 1996/33735; U.S. Pat. Nos.5,413,923; 5,625,126; 5,633,425; 5,569,825; 5,661,016; 5,545,896;5,814,318; 5,885,793; 5,916,771; and 5,939,598).

As used herein, the term “humanized” antibody refers to a non-humanantibody that contains minimal sequences derived from non-humanimmunoglobulin. In general, a humanized antibody contains substantiallyall of at least one, and typically two, variable domains, in which allor substantially all of the CDR regions correspond to those of anon-human immunoglobulin. All or substantially all of the FW regions mayalso be those of a human immunoglobulin sequence. The humanized antibodycan also contain at least a portion of an immunoglobulin constant region(Fc), typically that of a human immunoglobulin consensus sequence.Methods of antibody humanization are known in the art and have beendescribed, for example, in Riechmann et al. (1988) Nature 332:323-7;U.S. Pat. Nos. 5,530,101; 5,585,089; 5,693,761; 5,693,762; and6,180,370.

As used herein, patients that are “in need of” a hematopoietic stem celltransplant include patients that exhibit a defect or deficiency in oneor more blood cell types, as well as patients having a stem celldisorder, autoimmune disease, cancer, or other pathology describedherein. Hematopoietic stem cells generally exhibit 1) multi-potency, andcan thus differentiate into multiple different blood lineages including,but not limited to, granulocytes (e.g., promyelocytes, neutrophils,eosinophils, basophils), erythrocytes (e.g., reticulocytes,erythrocytes), thrombocytes (e.g., megakaryoblasts, platelet producingmegakaryocytes, platelets), monocytes (e.g., monocytes, macrophages),dendritic cells, microglia, osteoclasts, and lymphocytes (e.g., NKcells, B-cells and T-cells), 2) self-renewal, and can thus give rise todaughter cells that have equivalent potential as the mother cell, and 3)the ability to be reintroduced into a transplant recipient whereuponthey home to the hematopoietic stem cell niche and re-establishproductive and sustained hematopoiesis. Hematopoietic stem cells canthus be administered to a patient defective or deficient in one or morecell types of the hematopoietic lineage in order to re-constitute thedefective or deficient population of cells in vivo. For example, thepatient may be suffering from cancer, and the deficiency may be causedby administration of a chemotherapeutic agent or other medicament thatdepletes, either selectively or non-specifically, the cancerous cellpopulation. Additionally or alternatively, the patient may be sufferingfrom a hemoglobinopathy (e.g., a non-malignant hemoglobinopathy), suchas sickle cell anemia, thalassemia, Fanconi anemia, aplastic anemia, andWiskott-Aldrich syndrome. The subject may be one that is suffering fromadenosine deaminase severe combined immunodeficiency (ADA SCID),HIV/AIDS, metachromatic leukodystrophy, Diamond-Blackfan anemia, andSchwachman-Diamond syndrome. The subject may have or be affected by aninherited blood disorder (e.g., sickle cell anemia) or an autoimmunedisorder. Additionally or alternatively, the subject may have or beaffected by a malignancy, such as neuroblastoma or a hematologic cancer.In some embodiments, the subject may have a leukemia, lymphoma, ormyeloma. In some embodiments, the subject has acute myeloid leukemia,acute lymphoid leukemia, chronic myeloid leukemia, chronic lymphoidleukemia, multiple myeloma, diffuse large B-cell lymphoma, ornon-Hodgkin's lymphoma. In some embodiments, the subject hasmyelodysplastic syndrome. In some embodiments, the subject has anautoimmune disease, such as scleroderma, multiple sclerosis, ulcerativecolitis, Crohn's disease, Type 1 diabetes, or another autoimmunepathology described herein. In some embodiments, the subject is in needof chimeric antigen receptor T-cell (CART) therapy. In some embodiments,the subject has or is otherwise affected by a metabolic storagedisorder. The subject may suffer or otherwise be affected by a metabolicdisorder selected from the group consisting of glycogen storagediseases, mucopolysaccharidoses, Gaucher's Disease, Hurlers Disease,sphingolipidoses, metachromatic leukodystrophy, or any other diseases ordisorders which may benefit from the treatments and therapies disclosedherein and including, without limitation, severe combinedimmunodeficiency, Wiscott-Aldrich syndrome, hyper immunoglobulin M (IgM)syndrome, Chediak-Higashi disease, hereditary lymphohistiocytosis,osteopetrosis, osteogenesis imperfecta, storage diseases, thalassemiamajor, sickle cell disease, systemic sclerosis, systemic lupuserythematosus, multiple sclerosis, juvenile rheumatoid arthritis andthose diseases, or disorders described in “Bone Marrow Transplantationfor Non-Malignant Disease,” ASH Education Book, 1:319-338 (2000), thedisclosure of which is incorporated herein by reference in its entiretyas it pertains to pathologies that may be treated by administration ofhematopoietic stem cell transplant therapy. Additionally oralternatively, a patient “in need of” a hematopoietic stem celltransplant may one that is or is not suffering from one of the foregoingpathologies, but nonetheless exhibits a reduced level (e.g., as comparedto that of an otherwise healthy subject) of one or more endogenous celltypes within the hematopoietic lineage, such as megakaryocytes,thrombocytes, platelets, erythrocytes, mast cells, myeoblasts,basophils, neutrophils, eosinophils, microglia, granulocytes, monocytes,osteoclasts, antigen-presenting cells, macrophages, dendritic cells,natural killer cells, T-lymphocytes, and B-lymphocytes. One of skill inthe art can readily determine whether one's level of one or more of theforegoing cell types, or other blood cell type, is reduced with respectto an otherwise healthy subject, for example, by way of flow cytometryand fluorescence activated cell sorting (FACS) methods, among otherprocedures, known in the art.

As used herein, the term “leukocyte” refers to a heterogeneous group ofnucleated blood cell types, and excludes erythrocytes and platelets.Leukocytes can be divided into two general groups:polymorphonucleocytes, which include neutrophils, eosinophils, andbasophils, and mononucleocytes, which include lymphocytes and monocytes.Polymorphonucleocytes contain many cytoplasmic granules and a multilobednucleus and include the following: neutrophils, which are generallyamoeboid in shape, phagocytic, and stain with both basic and acidicdyes, and eosinophils and basophils, which contain cytoplasmic granulesthat stain with acidic dyes and with basic dyes, respectively.

As used herein, the term “lymphocyte” refers to a mononuclear leukocytethat is involved in the mounting of an immune response. In general,lymphocytes include B lymphocytes, T lymphocytes, and NK cells.

As used herein, the terms “mobilize” and “mobilization” refer toprocesses by which a population of hematopoietic stem or progenitorcells is released from a stem cell niche, such as the bone marrow of asubject, into circulation in the peripheral blood. Mobilization ofhematopoietic stem and progenitor cells can be monitored, for example,by assessing the quantity or concentration of hematopoietic stem orprogenitor cells in a peripheral blood sample isolated from a subject.For example, the peripheral blood sample may be withdrawn from thesubject, and the quantity or concentration of hematopoietic stem orprogenitor cells in the peripheral blood sample may subsequently beassessed, following the administration of a hematopoietic stem orprogenitor cell mobilization regimen to the subject. The mobilizationregimen may include, for example, a CXCR4 antagonist, such as a CXCR4antagonist described herein (e.g., plerixafor or a variant thereof), anda CXCR2 agonist, such as a CXCR2 agonist described herein (e.g., Gro-βor a variant thereof, such as a truncation of Gro-β, for example, Gro-βT). The quantity or concentration of hematopoietic stem or progenitorcells in the peripheral blood sample isolated from the subject followingadministration of the mobilization regimen may be compared to thequantity or concentration of hematopoietic stem or progenitor cells in aperipheral blood sample isolated from the subject prior toadministration of the mobilization regimen. An observation that thequantity or concentration of hematopoietic stem or progenitor cells hasincreased in the peripheral blood of the subject followingadministration of the mobilization regimen is an indication that thesubject is responding to the mobilization regimen, and thathematopoietic stem and progenitor cells have been released from one ormore stem cell niches, such as the bone marrow, into peripheral bloodcirculation. In some embodiments, an observation that the quantity orconcentration of hematopoietic stem or progenitor cells has increased inthe peripheral blood of the subject by 1%, 100%, 1,000%, or more (e.g.,by 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%,16%, 17%, 18%, 19%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%,70%, 75%, 80%, 85%, 90%, 95%, 100%, 200%, 300%, 400%, 500%, 600%, 700%,800%, 900%, 1,000%, or more) following administration of themobilization regimen is an indication that the subject is responding tothe mobilization regimen, and that hematopoietic stem and progenitorcells have been released from one or more stem cell niches, such as thebone marrow, into peripheral blood circulation. Methods for determiningthe quantity or concentration of hematopoietic stem or progenitor cellsare described herein and known in the art, and include, for example,flow cytometry techniques that quantify hematopoietic stem or progenitorcells on the basis of the antigen expression profile of such cells,which is described herein. For example, human HSCs are CD34⁺, CD38⁻,CD45RA⁻, CD90⁺, CD49F⁺, and lin− (negative for mature lineage markersincluding CD2, CD3, CD4, CD7, CD8, CD10, CD11B, CD19, CD20, CD56,CD235A). Additional methods for determining the quantity orconcentration of hematopoietic stem or progenitor cells in a peripheralblood sample isolated from a subject include assays that quantify thenumber of colony-forming units (CFUs) in the sample, which is a measureof the quantity of viable hematopoietic stem or progenitor cells that,upon incubation with an appropriate culture medium, give rise to anindividual population of hematopoietic stem or progenitor cells.

As used herein, the term “mobilizing amount” refers to a quantity of oneor more agents, such as a quantity of a CXCR4 antagonist and/or a CXCR2agonist described herein (In some embodiments, a quantity of plerixafor,or a variant thereof, and/or Gro-β, or a variant thereof, such as atruncation of Gro-β, for example, Gro-β T) that mobilizes a populationof hematopoietic stem or progenitor cells upon administration to asubject, such as a mammalian subject (e.g., a human subject). Exemplarymobilizing amounts of these agents include amounts sufficient toeffectuate the release of a population of, for example, from about 20 toabout 40 CD34⁺ cells/μL of peripheral blood, such as from about 21 toabout 39 CD34⁺ cells/μL of peripheral blood, about 22 to about 38 CD34⁺cells/4 of peripheral blood, about 23 to about 37 CD34⁺ cells/4 ofperipheral blood, about 24 to about 36 CD34⁺ cells/μL of peripheralblood, about 25 to about 35 CD34⁺ cells/4 of peripheral blood, about 26to about 34 CD34⁺ cells/4 of peripheral blood, about 27 to about 33CD34⁺ cells/4 of peripheral blood, about 28 to about 32 CD34⁺ cells/4 ofperipheral blood, or about 29 to about 31 CD34⁺ cells/4 of peripheralblood (e.g., about 20 CD34⁺ cells/4 of peripheral blood, 21 CD34⁺cells/4 of peripheral blood, 22 CD34⁺ cells/4 of peripheral blood, 23CD34⁺ cells/4 of peripheral blood, 24, CD34⁺ cells/μL of peripheralblood, 25 CD34⁺ cells/4 of peripheral blood, 26 CD34⁺ cells/4 ofperipheral blood, 27 CD34⁺ cells/4 of peripheral blood, 28 CD34⁺ cells/4of peripheral blood, 29 CD34⁺ cells/μL of peripheral blood, 30 CD34⁺cells/4 of peripheral blood, 31 CD34⁺ cells/4 of peripheral blood, 32CD34⁺ cells/4 of peripheral blood 33 CD34⁺ cells/4 of peripheral blood,34 CD34⁺ cells/4 of peripheral blood, 35 CD34⁺ cells/4 of peripheralblood, 36 CD34⁺ cells/4 of peripheral blood, 37 CD34⁺ cells/4 ofperipheral blood, 38 CD34⁺ cells/4 of peripheral blood, 39 CD34⁺ cells/4of peripheral blood, 40 CD34⁺ cells/μL of peripheral blood, or more. Forinstance, mobilizing amounts of a CXCR2 agonist, such as Gro-β T,include from about 50 μg/kg of recipient to about 1 mg/kg of recipient,such as from about 50 μg/kg to about 300 μg/kg, 100 μg/kg to about 250μg/kg, or about 150 μg/kg. Mobilizing amounts of a CXCR4 antagonist,such as plerixafor or a pharmaceutically acceptable salt thereof,include from about 50 μg/kg of recipient to about 500 μg/kg ofrecipient, such as from about 200 μg/kg to about 300 μg/kg, or about 240μg/kg.

As used herein, the term “monoclonal antibody” refers to an antibodythat is derived from a single clone, including any eukaryotic,prokaryotic, or phage clone, and not the method by which it is produced.

As used herein, the term “monocyte” refers to a CD14⁺ and CD34−peripheral blood mononuclear cell (PBMC), which is generally capable ofdifferentiating into a macrophage and/or dendritic cell upon activationby one or more foreign substances, such as, a microbial product. Inparticular, a monocyte may express elevated levels of the CD14 surfaceantigen marker, and may express at least one biomarker selected fromCD64, CD93, CD180, CD328 (also known as sialic acid-binding Ig-likelectin 7 or Siglec7), and CD329 (sialic acid-binding Ig-like lectin 9 orSiglec9), as well as the peanut agglutinin protein (PNA).

As used herein, a “peptide” refers to a single-chain polyamidecontaining a plurality of amino acid residues, such asnaturally-occurring and/or non-natural amino acid residues, that areconsecutively bound by amide bonds. Examples of peptides include shorterfragments of full-length proteins, such as full-lengthnaturally-occurring proteins.

As used herein, the term “recipient” refers to a patient that receives atransplant, such as a transplant containing a population ofhematopoietic stem cells. The transplanted cells administered to arecipient may be, e.g., autologous, syngeneic, or allogeneic cells.

As used herein, the term “sample” refers to a specimen (e.g., blood,blood component (e.g., serum or plasma), urine, saliva, amniotic fluid,cerebrospinal fluid, tissue (e.g., placental or dermal), pancreaticfluid, chorionic villus sample, and cells) taken from a subject. Asample may be, for example, withdrawn peripheral blood from a donor thatis undergoing or has undergone a hematopoietic stem or progenitor cellmobilization regimen described herein.

As used herein, the term “scFv” refers to a single chain Fv antibody inwhich the variable domains of the heavy chain and the light chain froman antibody have been joined to form one chain. scFv fragments contain asingle polypeptide chain that includes the variable region of anantibody light chain (V_(L)) (e.g., CDR-L1, CDR-L2, and/or CDR-L3) andthe variable region of an antibody heavy chain (V_(H)) (e.g., CDR-H1,CDR-H2, and/or CDR-H3) separated by a linker. The linker that joins theV_(L) and V_(H) regions of a scFv fragment can be a peptide linkercomposed of proteinogenic amino acids. Alternative linkers can be usedto so as to increase the resistance of the scFv fragment to proteolyticdegradation (for example, linkers containing D-amino acids), in order toenhance the solubility of the scFv fragment (for example, hydrophiliclinkers such as polyethylene glycol-containing linkers or polypeptidescontaining repeating glycine and serine residues), to improve thebiophysical stability of the molecule (for example, a linker containingcysteine residues that form intramolecular or intermolecular disulfidebonds), or to attenuate the immunogenicity of the scFv fragment (forexample, linkers containing glycosylation sites). It will also beunderstood by one of ordinary skill in the art that the variable regionsof the scFv molecules described herein can be modified such that theyvary in amino acid sequence from the antibody molecule from which theywere derived. For example, nucleotide or amino acid substitutionsleading to conservative substitutions or changes at amino acid residuescan be made (e.g., in CDR and/or framework residues) so as to preserveor enhance the ability of the scFv to bind to the antigen recognized bythe corresponding antibody.

As used herein, the phrase “stem cell disorder” broadly refers to anydisease, disorder, or condition that may be treated or cured byengrafting or transplanting a population of hematopoietic stem orprogenitor cells in a target tissue within a patient. For example, TypeI diabetes has been shown to be cured by hematopoietic stem celltransplant, along with various other disorders. Exemplary diseases thatcan be treated by infusion of hematopoietic stem or progenitor cellsinto a patient are sickle cell anemia, thalassemias, Fanconi anemia,aplastic anemia, Wiskott-Aldrich syndrome, ADA SCID, HIV/AIDS,metachromatic leukodystrophy, Diamond-Blackfan anemia, andSchwachman-Diamond syndrome. Additional diseases that may be treated bytransplantation of hematopoietic stem and progenitor cells as describedherein include blood disorders (e.g., sickle cell anemia) and autoimmunedisorders, such as scleroderma, multiple sclerosis, ulcerative colitis,and Crohn's disease. Additional diseases that may be treated usinghematopoietic stem and progenitor cell transplant therapy includecancer, such as a cancer described herein. Exemplary stem cell disordersare malignancies, such as a neuroblastoma or a hematologic cancers, suchas leukemia, lymphoma, and myeloma. In some embodiments, the cancer maybe acute myeloid leukemia, acute lymphoid leukemia, chronic myeloidleukemia, chronic lymphoid leukemia, multiple myeloma, diffuse largeB-cell lymphoma, or non-Hodgkin's lymphoma. Additional diseasestreatable using hematopoietic stem or progenitor cell transplant therapyinclude myelodysplastic syndrome. In some embodiments, the patient hasor is otherwise affected by a metabolic storage disorder. For example,the patient may suffer or otherwise be affected by a metabolic disorderselected from the group consisting of glycogen storage diseases,mucopolysaccharidoses, Gaucher's Disease, Hurlers Disease,sphingolipidoses, metachromatic leukodystrophy, or any other diseases ordisorders which may benefit from the treatments and therapies disclosedherein and including, without limitation, severe combinedimmunodeficiency, Wiscott-Aldrich syndrome, hyper immunoglobulin M (IgM)syndrome, Chediak-Higashi disease, hereditary lymphohistiocytosis,osteopetrosis, osteogenesis imperfecta, storage diseases, thalassemiamajor, sickle cell disease, systemic sclerosis, systemic lupuserythematosus, multiple sclerosis, juvenile rheumatoid arthritis andthose diseases, or disorders described in “Bone Marrow Transplantationfor Non-Malignant Disease,” ASH Education Book, 1:319-338 (2000), thedisclosure of which is incorporated herein by reference in its entiretyas it pertains to pathologies that may be treated by administration ofhematopoietic stem or progenitor cell transplant therapy.

As used herein in the context of hematopoietic stem cell mobilization,the term “stem cell niche” refers to a microenvironment within a donor,such as a mammalian donor (e.g., a human donor) in which endogenoushematopoietic stem or progenitor cells reside. An exemplary stem cellniche is bone marrow tissue.

As used herein, the terms “subject” and “patient” refer to an organism,such as a human, that receives treatment for a particular disease orcondition as described herein. In some embodiments, a patient, such as ahuman patient, that is in need of hematopoietic stem celltransplantation may receive treatment that includes a population ofhematopoietic stem cells so as to treat a stem cell disorder, such as acancer, autoimmune disease, or metabolic disorder described herein. Thehematopoietic stem cells that are transplanted into the patient may be,for example, a population of hematopoietic stem cells that has beenmobilized and withdrawn from a donor in accordance with the compositionsand methods described herein. In some embodiments, the hematopoieticstem cells that are transplanted into the patient may be mobilizedwithin a donor by administration of a CXCR4 antagonist and/or a CXCR2agonist to the donor.

As used herein, the term “transfection” refers to any of a wide varietyof techniques commonly used for the introduction of exogenous DNA into aprokaryotic or eukaryotic host cell, such as electroporation,lipofection, calcium-phosphate precipitation, DEAE-dextran transfectionand the like.

As used herein, the terms “treat” or “treatment” refer to therapeutictreatment, in which the object is to prevent or slow down (lessen) anundesired physiological change or disorder or to promote a beneficialphenotype in the patient being treated. Beneficial or desired clinicalresults include, but are not limited to, promoting the engraftment ofexogenous hematopoietic cells in a patient following hematopoietic stemor progenitor cell transplant therapy. In certain embodiments, thebenefits include a more rapid engraftment of transplanted cells, e.g.,neutrophils and platelets. For example, in certain embodiments, usingthe methods described herein, neutrophil recovery occurs within about5-20 days post-transplant, about 5-15 days post-transplant, about 5-10days post-transplant, about 7-12 days post-transplant, about 8-12 dayspost transplant, about 9-15 days post-transplant, about 10-15 dayspost-transplant, or about 10 days post-transplant. In certainembodiments, using the methods described herein, platelet recoveryoccurs within about 10-20 days post-transplant, about 10-15 dayspost-transplant, about 15-20 days post-transplant, about 12-18 dayspost-transplant, about 12-17 days post transplant, about 13-18 dayspost-transplant, about 12-17 days post-transplant, or about 15 dayspost-transplant. Additional beneficial results include an increase inthe cell count or relative concentration of hematopoietic stem cells ina patient in need of a hematopoietic stem or progenitor cell transplantfollowing administration of an exogenous hematopoietic stem orprogenitor cell graft to the patient. Beneficial results of therapydescribed herein may also include an increase in the cell count orrelative concentration of one or more cells of hematopoietic lineage,such as a megakaryocyte, thrombocyte, platelet, erythrocyte, mast cell,myeoblast, basophil, neutrophil, eosinophil, microglial cell,granulocyte, monocyte, osteoclast, antigen-presenting cell, macrophage,dendritic cell, natural killer cell, T-lymphocyte, or B-lymphocyte,following and subsequent hematopoietic stem cell transplant therapy.Additional beneficial results may include the reduction in quantity of adisease-causing cell population, such as a population of cancer cells orautoimmune cells.

As used herein, the terms “variant” and “derivative” are usedinterchangeably and refer to naturally-occurring, synthetic, andsemi-synthetic analogues of a compound, peptide, protein, or othersubstance described herein. A variant or derivative of a compound,peptide, protein, or other substance described herein may retain orimprove upon the biological activity of the original material.

As used herein, the term “vector” includes a nucleic acid vector, suchas a plasmid, a DNA vector, a plasmid, a RNA vector, virus, or othersuitable replicon. Expression vectors described herein may contain apolynucleotide sequence as well as, for example, additional sequenceelements used for the expression of proteins and/or the integration ofthese polynucleotide sequences into the genome of a mammalian cell.Certain vectors that can be used for the expression of peptides andproteins, such as those described herein, include plasmids that containregulatory sequences, such as promoter and enhancer regions, whichdirect gene transcription. Other useful vectors for expression ofpeptides and proteins described herein contain polynucleotide sequencesthat enhance the rate of translation of these genes or improve thestability or nuclear export of the mRNA that results from genetranscription. These sequence elements may include, for example, 5′ and3′ untranslated regions and a polyadenylation signal site in order todirect efficient transcription of the gene carried on the expressionvector. The expression vectors described herein may also contain apolynucleotide encoding a marker for selection of cells that containsuch a vector. Examples of a suitable marker include genes that encoderesistance to antibiotics, such as ampicillin, chloramphenicol,kanamycin, and nourseothricin.

As used herein, the term “alkyl” refers to a straight- or branched-chainalkyl group having, for example, from 1 to 20 carbon atoms in the chain.Examples of alkyl groups include methyl, ethyl, n-propyl, isopropyl,butyl, isobutyl, sec-butyl, tert-butyl, pentyl, isopentyl, tert-pentyl,hexyl, isohexyl, and the like.

As used herein, the term “alkylene” refers to a straight- orbranched-chain divalent alkyl group. The divalent positions may be onthe same or different atoms within the alkyl chain. Examples of alkyleneinclude methylene, ethylene, propylene, isopropylene, and the like.

As used herein, the term “heteroalkyl” refers to a straight orbranched-chain alkyl group having, for example, from 1 to 20 carbonatoms in the chain, and further containing one or more heteroatoms(e.g., oxygen, nitrogen, or sulfur, among others) in the chain.

As used herein, the term “heteroalkylene” refers to a straight- orbranched-chain divalent heteroalkyl group. The divalent positions may beon the same or different atoms within the heteroalkyl chain. Thedivalent positions may be one or more heteroatoms.

As used herein, the term “alkenyl” refers to a straight- orbranched-chain alkenyl group having, for example, from 2 to 20 carbonatoms in the chain. Examples of alkenyl groups include vinyl, propenyl,isopropenyl, butenyl, tert-butylenyl, hexenyl, and the like.

As used herein, the term “alkenylene” refers to a straight- orbranched-chain divalent alkenyl group. The divalent positions may be onthe same or different atoms within the alkenyl chain. Examples ofalkenylene include ethenylene, propenylene, isopropenylene, butenylene,and the like.

As used herein, the term “heteroalkenyl” refers to a straight- orbranched-chain alkenyl group having, for example, from 2 to 20 carbonatoms in the chain, and further containing one or more heteroatoms(e.g., oxygen, nitrogen, or sulfur, among others) in the chain.

As used herein, the term “heteroalkenylene” refers to a straight- orbranched-chain divalent heteroalkenyl group. The divalent positions maybe on the same or different atoms within the heteroalkenyl chain. Thedivalent positions may be one or more heteroatoms.

As used herein, the term “alkynyl” refers to a straight- orbranched-chain alkynyl group having, for example, from 2 to 20 carbonatoms in the chain. Examples of alkynyl groups include propargyl,butynyl, pentynyl, hexynyl, and the like.

As used herein, the term “alkynylene” refers to a straight- orbranched-chain divalent alkynyl group. The divalent positions may be onthe same or different atoms within the alkynyl chain.

As used herein, the term “heteroalkynyl” refers to a straight- orbranched-chain alkynyl group having, for example, from 2 to 20 carbonatoms in the chain, and further containing one or more heteroatoms(e.g., oxygen, nitrogen, or sulfur, among others) in the chain.

As used herein, the term “heteroalkynylene” refers to a straight- orbranched-chain divalent heteroalkynyl group. The divalent positions maybe on the same or different atoms within the heteroalkynyl chain. Thedivalent positions may be one or more heteroatoms.

As used herein, the term “cycloalkyl” refers to a monocyclic, or fused,bridged, or spiro polycyclic ring structure that is saturated and has,for example, from 3 to 12 carbon ring atoms. Examples of cycloalkylgroups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,cycloheptyl, cyclooctyl, bicyclo[3.1.0]hexane, and the like.

As used herein, the term “cycloalkylene” refers to a divalent cycloalkylgroup. The divalent positions may be on the same or different atomswithin the ring structure. Examples of cycloalkylene includecyclopropylene, cyclobutylene, cyclopentylene, cyclohexylene, and thelike.

As used herein, the term “heterocycloalkyl” refers to a monocyclic, orfused, bridged, or spiro polycyclic ring structure that is saturated andhas, for example, from 3 to 12 ring atoms per ring structure selectedfrom carbon atoms and heteroatoms selected from, e.g., nitrogen, oxygen,and sulfur, among others. The ring structure may contain, for example,one or more oxo groups on carbon, nitrogen, or sulfur ring members.

As used herein, the term “heterocycloalkylene” refers to a divalentheterocyclolalkyl group. The divalent positions may be on the same ordifferent atoms within the ring structure.

As used herein, the term “aryl” refers to a monocyclic or multicyclicaromatic ring system containing, for example, from 6 to 19 carbon atoms.Aryl groups include, but are not limited to, phenyl, fluorenyl,naphthyl, and the like. The divalent positions may be one or moreheteroatoms.

As used herein, the term “arylene” refers to a divalent aryl group. Thedivalent positions may be on the same or different atoms.

As used herein, the term “heteroaryl” refers to a monocyclicheteroaromatic, or a bicyclic or a tricyclic fused-ring heteroaromaticgroup. Heteroaryl groups include pyridyl, pyrrolyl, furyl, thienyl,imidazolyl, oxazolyl, isoxazolyl, thiazolyl, isothiazolyl, pyrazolyl,1,2,3-triazolyl, 1,2,4-triazolyl, 1,2,3-oxadiazolyl, 1,2,4-oxadia-zolyl,1,2,5-oxadiazolyl, 1,3,4-oxadiazolyl, 1,3,4-triazinyl, 1,2,3-triazinyl,benzofuryl, [2,3-dihydro]benzofuryl, isobenzofuryl, benzothienyl,benzotriazolyl, isobenzothienyl, indolyl, isoindolyl, 3H-indolyl,benzimidazolyl, imidazo[1,2-a]pyridyl, benzothiazolyl, benzoxazolyl,quinolizinyl, quinazolinyl, phthalazinyl, quinoxalinyl, cinnolinyl,napthyridinyl, pyrido[3,4-b]pyridyl, pyrido[3,2-b]pyridyl,pyrido[4,3-b]pyridyl, quinolyl, isoquinolyl, tetrazolyl,5,6,7,8-tetrahydroquinolyl, 5,6,7,8-tetrahydroisoquinolyl, purinyl,pteridinyl, carbazolyl, xanthenyl, benzoquinolyl, and the like.

As used herein, the term “heteroarylene” refers to a divalent heteroarylgroup. The divalent positions may be on the same or different atoms. Thedivalent positions may be one or more heteroatoms.

Unless otherwise constrained by the definition of the individualsubstituent, the foregoing chemical moieties, such as “alkyl”,“alkylene”, “heteroalkyl”, “heteroalkylene”, “alkenyl”, “alkenylene”,“heteroalkenyl”, “heteroalkenylene”, “alkynyl”, “alkynylene”,“heteroalkynyl”, “heteroalkynylene”, “cycloalkyl”, “cycloalkylene”,“heterocyclolalkyl”, heterocycloalkylene”, “aryl,” “arylene”,“heteroaryl”, and “heteroarylene” groups can optionally be substituted.As used herein, the term “optionally substituted” refers to a compoundor moiety containing one or more (for example, 1, 2, 3, 4, 5, 6, 7, 8,9, 10, or more) substituents, as permitted by the valence of thecompound or moiety or a site thereof, such as a substituent selectedfrom the group consisting of alkyl, alkenyl, alkynyl, cycloalkyl,heterocycloalkyl, alkyl aryl, alkyl heteroaryl, alkyl cycloalkyl, alkylheterocycloalkyl, amino, ammonium, acyl, acyloxy, acylamino,aminocarbonyl, alkoxycarbonyl, ureido, carbamate, aryl, heteroaryl,sulfinyl, sulfonyl, alkoxy, sulfanyl, halogen, carboxy, trihalomethyl,cyano, hydroxy, mercapto, nitro, and the like. The substitution mayinclude situations in which neighboring substituents have undergone ringclosure, such as ring closure of vicinal functional substituents, toform, for example, lactams, lactones, cyclic anhydrides, acetals,hemiacetals, thioacetals, aminals, and hemiaminals, formed by ringclosure, for example, to furnish a protecting group.

Methods of Mobilizing Hematopoietic Stem and Progenitor Cells andReleasing Cells for Expansion and Therapeutic Use

The present invention is based, in part, on the discovery thathematopoietic stem and progenitor cells can be mobilized byadministering particular doses of a CXCR2 agonist, such as Gro-β, Gro-βT, or a variant thereof, optionally in combination with a CXCR4antagonist to a mammalian donor (e.g., a human donor) while reducing themobilization of other cell types, such as leukocytes, neutrophils,lymphocytes, and monocytes. This property is particularly beneficial inthe context of hematopoietic stem cell transplant therapy, ashematopoietic stem cells that are mobilized and isolated from a donorusing the compositions and method described herein have reducedquantities of cell types that are undesirable for administration to ahuman patient suffering from a stem cell disorder.

Particularly, it has been discovered that CXCR2 agonists, such as Gro-β,Gro-β T, or a variant thereof, when administered intravenously at a doseof from about 50 μg/kg to about 1 mg/kg, preferably from about 100 μg/kgto about 250 μg/kg, and even more preferably at a dose of about 150μg/kg, exhibit the ability to rapidly mobilize hematopoietic stem andprogenitor cells in a donor (e.g., a mammalian donor, such as a humandonor) while reducing the mobilization of other cells of thehematopoietic lineage that may be undesirable for infusion into apatient (e.g., a mammalian patient, such as a human patient) that isundergoing hematopoietic stem cell transplant therapy. CXCR2 agonists,such as Gro-β, Gro-β T, or a variant thereof, when administered at theabove doses to a donor exhibit the ability to selectively mobilizehematopoietic stem cells as described in detail in Example 1, below.

When determining whether hematopoietic stem cells mobilized in a donorby administration of a CXCR2 agonist, such as Gro-β, Gro-β T, or avariant thereof, and optionally, a CXCR4 antagonist, such as plerixaforor a pharmaceutically acceptable salt thereof, are suitable for releasefor ex vivo expansion and/or for therapeutic use, one may acquire aninput value for each of one or more parameters set forth in TABLE 2 thatcharacterize a sample of peripheral blood of the donor. The one or moreparameters may be compared to the corresponding reference criterion foreach parameter, and if the reference criterion is satisfied by the ampleof hematopoietic stem cells, the cells isolated from the donor may bereleased for expansion ex vivo and/or for infusion into a patient fortherapeutic use (e.g., for the treatment of one or more stem celldisorders described herein).

Exemplary hematopoietic stem cell parameters and corresponding referencecriteria useful in conjunction with the compositions and methodsdescribed herein are set forth in TABLE 2, below.

TABLE 2 Hematopoietic stem cell population parameters and correspondingreference criteria Parameter Parameter Category No. Parameter ReferenceCriterion Ratio of CD34⁺ cells 1 Ratio of CD34⁺ cells to At least 0.0006(e.g., CD34⁺ CD90⁺ leukocytes CD45RA⁻ cells) to 2 Ratio of CD34⁺ cellsto At least 0.0009 other cells of the leukocytes hematopoietic 3 Ratioof CD34⁺ CD90⁺ At least 0.0002 lineage within CD45RA⁻ cells toleukocytes peripheral blood 4 Ratio of CD34⁺ CD90⁺ At least 0.0003sample isolated from CD45RA⁻ cells to leukocytes mammalian donor 5 Ratioof CD34⁺ CD90⁺ At least 0.0004 CD45RA⁻ cells to leukocytes 6 Ratio ofCD34⁺ cells to At least 0.0011 neutrophils 7 Ratio of CD34⁺ cells to Atleast 0.0004 neutrophils 8 Ratio of CD34⁺ CD90⁺ At least 0.0006 CD45RA⁻cells to neutrophils 9 Ratio of CD34⁺ CD90⁺ At least 0.0007 CD45RA⁻cells to neutrophils 10 Ratio of CD34⁺ cells to At least 0.0020lymphocytes 11 Ratio of CD34⁺ cells to At least 0.0025 lymphocytes 12Ratio of CD34⁺ CD90⁺ At least 0.0005 CD45RA⁻ cells to lymphocytes 13Ratio of CD34⁺ CD90⁺ At least 0.0011 CD45RA⁻ cells to lymphocytes 14Ratio of CD34⁺ cells to At least 0.0047 monocytes 15 Ratio of CD34⁺cells to At least 0.0111 monocytes 16 Ratio of CD34⁺ CD90⁺ At least0.0020 CD45RA⁻ cells to monocytes 17 Ratio of CD34⁺ CD90⁺ At least0.0039 CD45RA⁻ cells to monocytes Proportion of CD34⁺ 18 Frequency ofCD34⁺ cells At least 0.051% cells (e.g., CD34⁺ relative to totalquantity of CD90⁺ CD45RA⁻ cells in sample isolated from cells) inperipheral donor blood sample isolated 19 Frequency of CD34⁺ cells Atleast 0.097% from donor relative to total quantity of cells in sampleisolated from donor 20 Frequency of CD34⁺ CD90⁺ At least 0.020% CD45RA⁻cells relative to total quantity of cells in sample isolated from donor21 Frequency of CD34⁺ CD90⁺ At least 0.066% CD45RA⁻ cells relative tototal quantity of cells in sample isolated from donor

In selecting parameters for determining whether a population ofhematopoietic stem cells obtained from a donor (e.g., a mammalian donor,such as a human donor) is suitable for release for ex vivo expansion ortherapeutic use, one may select one or more input parameters listed inTABLE 2. In some embodiments, one may select an individual parameterfrom parameter numbers 1-21. Alternatively, one may select a combinationof parameters, such as a CD34⁺ cell ratio parameter (e.g., one or moreof parameter numbers 1-17 in TABLE 2) and a frequency parameter (e.g.,one or more of parameter numbers 18-21 listed in TABLE 2). In someembodiments, the parameters used for determining whether a population ofhematopoietic stem cells obtained from a donor (e.g., a mammalian donor,such as a human donor) is suitable for release for ex vivo expansion ortherapeutic use are a combination of parameters as set forth in any oneof TABLES 3-6, below.

TABLE 3 Two-way combinations of hematopoietic stem cell populationparameters for assessment First Parameter Second No. Parameter No. 1 6 17 1 8 1 9 2 6 2 7 2 8 2 9 3 6 3 7 3 8 3 9 4 6 4 7 4 8 4 9 5 6 5 7 5 8 59 1 10 1 11 1 12 1 13 2 10 2 11 2 12 2 13 3 10 3 11 3 12 3 13 4 10 4 114 12 4 13 5 10 5 11 5 12 5 13 1 14 1 15 1 16 1 17 2 14 2 15 2 16 2 17 314 3 15 3 16 3 17 4 14 4 15 4 16 4 17 5 14 5 15 5 16 5 17 1 18 1 19 1 201 21 2 18 2 19 2 20 2 21 3 18 3 19 3 20 3 21 4 18 4 19 4 20 4 21 5 18 519 5 20 5 21 6 10 6 11 6 12 6 13 7 10 7 11 7 12 7 13 8 10 8 11 8 12 8 139 10 9 11 9 12 9 13 6 14 6 15 6 16 6 17 7 14 7 15 7 16 7 17 8 14 8 15 816 8 17 9 14 9 15 9 16 9 17 6 18 6 19 6 20 6 21 7 18 7 19 7 20 7 21 8 188 19 8 20 8 21 9 18 9 19 9 20 9 21 10 14 10 15 10 16 10 17 11 14 11 1511 16 11 17 12 14 12 15 12 16 12 17 13 14 13 15 13 16 13 17 10 18 10 1910 20 10 21 11 18 11 19 11 20 11 21 12 18 12 19 12 20 12 21 13 18 13 1913 20 13 21 14 18 14 19 14 20 14 21 15 18 15 19 15 20 15 21 16 18 16 1916 20 16 21 17 18 17 19 17 20 17 21

TABLE 4 Three-way combinations of hematopoietic stem cell populationparameters for assessment First Parameter Second Parameter ThirdParameter No. No. No. 1 6 10 1 7 10 1 8 10 1 9 10 2 6 10 2 7 10 2 8 10 29 10 3 6 10 3 7 10 3 8 10 3 9 10 4 6 10 4 7 10 4 8 10 4 9 10 5 6 10 5 710 5 8 10 5 9 10 1 6 11 1 7 11 1 8 11 1 9 11 2 6 11 2 7 11 2 8 11 2 9 113 6 11 3 7 11 3 8 11 3 9 11 4 6 11 4 7 11 4 8 11 4 9 11 5 6 11 5 7 11 58 11 5 9 11 1 6 12 1 7 12 1 8 12 1 9 12 2 6 12 2 7 12 2 8 12 2 9 12 3 612 3 7 12 3 8 12 3 9 12 4 6 12 4 7 12 4 8 12 4 9 12 5 6 12 5 7 12 5 8 125 9 12 1 6 13 1 7 13 1 8 13 1 9 13 2 6 13 2 7 13 2 8 13 2 9 13 3 6 13 37 13 3 8 13 3 9 13 4 6 13 4 7 13 4 8 13 4 9 13 5 6 13 5 7 13 5 8 13 5 913 1 6 14 1 7 14 1 8 14 1 9 14 2 6 14 2 7 14 2 8 14 2 9 14 3 6 14 3 7 143 8 14 3 9 14 4 6 14 4 7 14 4 8 14 4 9 14 5 6 14 5 7 14 5 8 14 5 9 14 16 15 1 7 15 1 8 15 1 9 15 2 6 15 2 7 15 2 8 15 2 9 15 3 6 15 3 7 15 3 815 3 9 15 4 6 15 4 7 15 4 8 15 4 9 15 5 6 15 5 7 15 5 8 15 5 9 15 1 6 161 7 16 1 8 16 1 9 16 2 6 16 2 7 16 2 8 16 2 9 16 3 6 16 3 7 16 3 8 16 39 16 4 6 16 4 7 16 4 8 16 4 9 16 5 6 16 5 7 16 5 8 16 5 9 16 1 6 17 1 717 1 8 17 1 9 17 2 6 17 2 7 17 2 8 17 2 9 17 3 6 17 3 7 17 3 8 17 3 9 174 6 17 4 7 17 4 8 17 4 9 17 5 6 17 5 7 17 5 8 17 5 9 17 1 6 18 1 7 18 18 18 1 9 18 2 6 18 2 7 18 2 8 18 2 9 18 3 6 18 3 7 18 3 8 18 3 9 18 4 618 4 7 18 4 8 18 4 9 18 5 6 18 5 7 18 5 8 18 5 9 18 1 6 19 1 7 19 1 8 191 9 19 2 6 19 2 7 19 2 8 19 2 9 19 3 6 19 3 7 19 3 8 19 3 9 19 4 6 19 47 19 4 8 19 4 9 19 5 6 19 5 7 19 5 8 19 5 9 19 1 6 20 1 7 20 1 8 20 1 920 2 6 20 2 7 20 2 8 20 2 9 20 3 6 20 3 7 20 3 8 20 3 9 20 4 6 20 4 7 204 8 20 4 9 20 5 6 20 5 7 20 5 8 20 5 9 20 1 6 21 1 7 21 1 8 21 1 9 21 26 21 2 7 21 2 8 21 2 9 21 3 6 21 3 7 21 3 8 21 3 9 21 4 6 21 4 7 21 4 821 4 9 21 5 6 21 5 7 21 5 8 21 5 9 21 1 10 14 1 11 14 1 12 14 1 13 14 210 14 2 11 14 2 12 14 2 13 14 3 10 14 3 11 14 3 12 14 3 13 14 4 10 14 411 14 4 12 14 4 13 14 5 10 14 5 11 14 5 12 14 5 13 14 1 10 15 1 11 15 112 15 1 13 15 2 10 15 2 11 15 2 12 15 2 13 15 3 10 15 3 11 15 3 12 15 313 15 4 10 15 4 11 15 4 12 15 4 13 15 5 10 15 5 11 15 5 12 15 5 13 15 110 16 1 11 16 1 12 16 1 13 16 2 10 16 2 11 16 2 12 16 2 13 16 3 10 16 311 16 3 12 16 3 13 16 4 10 16 4 11 16 4 12 16 4 13 16 5 10 16 5 11 16 512 16 5 13 16 1 10 17 1 11 17 1 12 17 1 13 17 2 10 17 2 11 17 2 12 17 213 17 3 10 17 3 11 17 3 12 17 3 13 17 4 10 17 4 11 17 4 12 17 4 13 17 510 17 5 11 17 5 12 17 5 13 17 1 10 18 1 11 18 1 12 18 1 13 18 2 10 18 211 18 2 12 18 2 13 18 3 10 18 3 11 18 3 12 18 3 13 18 4 10 18 4 11 18 412 18 4 13 18 5 10 18 5 11 18 5 12 18 5 13 18 1 10 19 1 11 19 1 12 19 113 19 2 10 19 2 11 19 2 12 19 2 13 19 3 10 19 3 11 19 3 12 19 3 13 19 410 19 4 11 19 4 12 19 4 13 19 5 10 19 5 11 19 5 12 19 5 13 19 1 10 20 111 20 1 12 20 1 13 20 2 10 20 2 11 20 2 12 20 2 13 20 3 10 20 3 11 20 312 20 3 13 20 4 10 20 4 11 20 4 12 20 4 13 20 5 10 20 5 11 20 5 12 20 513 20 1 10 21 1 11 21 1 12 21 1 13 21 2 10 21 2 11 21 2 12 21 2 13 21 310 21 3 11 21 3 12 21 3 13 21 4 10 21 4 11 21 4 12 21 4 13 21 5 10 21 511 21 5 12 21 5 13 21 1 14 18 1 15 18 1 16 18 1 17 18 2 14 18 2 15 18 216 18 2 17 18 3 14 18 3 15 18 3 16 18 3 17 18 4 14 18 4 15 18 4 16 18 417 18 5 14 18 5 15 18 5 16 18 5 17 18 1 14 19 1 15 19 1 16 19 1 17 19 214 19 2 15 19 2 16 19 2 17 19 3 14 19 3 15 19 3 16 19 3 17 19 4 14 19 415 19 4 16 19 4 17 19 5 14 19 5 15 19 5 16 19 5 17 19 1 14 20 1 15 20 116 20 1 17 20 2 14 20 2 15 20 2 16 20 2 17 20 3 14 20 3 15 20 3 16 20 317 20 4 14 20 4 15 20 4 16 20 4 17 20 5 14 20 5 15 20 5 16 20 5 17 20 114 21 1 15 21 1 16 21 1 17 21 2 14 21 2 15 21 2 16 21 2 17 21 3 14 21 315 21 3 16 21 3 17 21 4 14 21 4 15 21 4 16 21 4 17 21 5 14 21 5 15 21 516 21 5 17 21 6 10 14 6 11 14 6 12 14 6 13 14 7 10 14 7 11 14 7 12 14 713 14 8 10 14 8 11 14 8 12 14 8 13 14 9 10 14 9 11 14 9 12 14 9 13 14 610 15 6 11 15 6 12 15 6 13 15 7 10 15 7 11 15 7 12 15 7 13 15 8 10 15 811 15 8 12 15 8 13 15 9 10 15 9 11 15 9 12 15 9 13 15 6 10 16 6 11 16 612 16 6 13 16 7 10 16 7 11 16 7 12 16 7 13 16 8 10 16 8 11 16 8 12 16 813 16 9 10 16 9 11 16 9 12 16 9 13 16 6 10 17 6 11 17 6 12 17 6 13 17 710 17 7 11 17 7 12 17 7 13 17 8 10 17 8 11 17 8 12 17 8 13 17 9 10 17 911 17 9 12 17 9 13 17 6 10 18 6 11 18 6 12 18 6 13 18 7 10 18 7 11 18 712 18 7 13 18 8 10 18 8 11 18 8 12 18 8 13 18 9 10 18 9 11 18 9 12 18 913 18 6 10 19 6 11 19 6 12 19 6 13 19 7 10 19 7 11 19 7 12 19 7 13 19 810 19 8 11 19 8 12 19 8 13 19 9 10 19 9 11 19 9 12 19 9 13 19 6 10 20 611 20 6 12 20 6 13 20 7 10 20 7 11 20 7 12 20 7 13 20 8 10 20 8 11 20 812 20 8 13 20 9 10 20 9 11 20 9 12 20 9 13 20 6 10 21 6 11 21 6 12 21 613 21 7 10 21 7 11 21 7 12 21 7 13 21 8 10 21 8 11 21 8 12 21 8 13 21 910 21 9 11 21 9 12 21 9 13 21 6 14 18 6 15 18 6 16 18 6 17 18 7 14 18 715 18 7 16 18 7 17 18 8 14 18 8 15 18 8 16 18 8 17 18 9 14 18 9 15 18 916 18 9 17 18 6 14 19 6 15 19 6 16 19 6 17 19 7 14 19 7 15 19 7 16 19 717 19 8 14 19 8 15 19 8 16 19 8 17 19 9 14 19 9 15 19 9 16 19 9 17 19 614 20 6 15 20 6 16 20 6 17 20 7 14 20 7 15 20 7 16 20 7 17 20 8 14 20 815 20 8 16 20 8 17 20 9 14 20 9 15 20 9 16 20 9 17 20 6 14 21 6 15 21 616 21 6 17 21 7 14 21 7 15 21 7 16 21 7 17 21 8 14 21 8 15 21 8 16 21 817 21 9 14 21 9 15 21 9 16 21 9 17 21 10 14 18 10 15 18 10 16 18 10 1718 11 14 18 11 15 18 11 16 18 11 17 18 12 14 18 12 15 18 12 16 18 12 1718 13 14 18 13 15 18 13 16 18 13 17 18 10 14 19 10 15 19 10 16 19 10 1719 11 14 19 11 15 19 11 16 19 11 17 19 12 14 19 12 15 19 12 16 19 12 1719 13 14 19 13 15 19 13 16 19 13 17 19 10 14 20 10 15 20 10 16 20 10 1720 11 14 20 11 15 20 11 16 20 11 17 20 12 14 20 12 15 20 12 16 20 12 1720 13 14 20 13 15 20 13 16 20 13 17 20 10 14 21 10 15 21 10 16 21 10 1721 11 14 21 11 15 21 11 16 21 11 17 21 12 14 21 12 15 21 12 16 21 12 1721 13 14 21 13 15 21 13 16 21 13 17 21

TABLE 5 Four-way combinations of hematopoietic stem cell populationparameters for assessment First Parameter Second Third Parameter FourthNo. Parameter No. No. Parameter No. 1 6 10 14 1 7 10 14 1 8 10 14 1 9 1014 2 6 10 14 2 7 10 14 2 8 10 14 2 9 10 14 3 6 10 14 3 7 10 14 3 8 10 143 9 10 14 4 6 10 14 4 7 10 14 4 8 10 14 4 9 10 14 5 6 10 14 5 7 10 14 58 10 14 5 9 10 14 1 6 11 14 1 7 11 14 1 8 11 14 1 9 11 14 2 6 11 14 2 711 14 2 8 11 14 2 9 11 14 3 6 11 14 3 7 11 14 3 8 11 14 3 9 11 14 4 6 1114 4 7 11 14 4 8 11 14 4 9 11 14 5 6 11 14 5 7 11 14 5 8 11 14 5 9 11 141 6 12 14 1 7 12 14 1 8 12 14 1 9 12 14 2 6 12 14 2 7 12 14 2 8 12 14 29 12 14 3 6 12 14 3 7 12 14 3 8 12 14 3 9 12 14 4 6 12 14 4 7 12 14 4 812 14 4 9 12 14 5 6 12 14 5 7 12 14 5 8 12 14 5 9 12 14 1 6 13 14 1 7 1314 1 8 13 14 1 9 13 14 2 6 13 14 2 7 13 14 2 8 13 14 2 9 13 14 3 6 13 143 7 13 14 3 8 13 14 3 9 13 14 4 6 13 14 4 7 13 14 4 8 13 14 4 9 13 14 56 13 14 5 7 13 14 5 8 13 14 5 9 13 14 1 6 10 15 1 7 10 15 1 8 10 15 1 910 15 2 6 10 15 2 7 10 15 2 8 10 15 2 9 10 15 3 6 10 15 3 7 10 15 3 8 1015 3 9 10 15 4 6 10 15 4 7 10 15 4 8 10 15 4 9 10 15 5 6 10 15 5 7 10 155 8 10 15 5 9 10 15 1 6 11 15 1 7 11 15 1 8 11 15 1 9 11 15 2 6 11 15 27 11 15 2 8 11 15 2 9 11 15 3 6 11 15 3 7 11 15 3 8 11 15 3 9 11 15 4 611 15 4 7 11 15 4 8 11 15 4 9 11 15 5 6 11 15 5 7 11 15 5 8 11 15 5 9 1115 1 6 12 15 1 7 12 15 1 8 12 15 1 9 12 15 2 6 12 15 2 7 12 15 2 8 12 152 9 12 15 3 6 12 15 3 7 12 15 3 8 12 15 3 9 12 15 4 6 12 15 4 7 12 15 48 12 15 4 9 12 15 5 6 12 15 5 7 12 15 5 8 12 15 5 9 12 15 1 6 13 15 1 713 15 1 8 13 15 1 9 13 15 2 6 13 15 2 7 13 15 2 8 13 15 2 9 13 15 3 6 1315 3 7 13 15 3 8 13 15 3 9 13 15 4 6 13 15 4 7 13 15 4 8 13 15 4 9 13 155 6 13 15 5 7 13 15 5 8 13 15 5 9 13 15 1 6 10 16 1 7 10 16 1 8 10 16 19 10 16 2 6 10 16 2 7 10 16 2 8 10 16 2 9 10 16 3 6 10 16 3 7 10 16 3 810 16 3 9 10 16 4 6 10 16 4 7 10 16 4 8 10 16 4 9 10 16 5 6 10 16 5 7 1016 5 8 10 16 5 9 10 16 1 6 11 16 1 7 11 16 1 8 11 16 1 9 11 16 2 6 11 162 7 11 16 2 8 11 16 2 9 11 16 3 6 11 16 3 7 11 16 3 8 11 16 3 9 11 16 46 11 16 4 7 11 16 4 8 11 16 4 9 11 16 5 6 11 16 5 7 11 16 5 8 11 16 5 911 16 1 6 12 16 1 7 12 16 1 8 12 16 1 9 12 16 2 6 12 16 2 7 12 16 2 8 1216 2 9 12 16 3 6 12 16 3 7 12 16 3 8 12 16 3 9 12 16 4 6 12 16 4 7 12 164 8 12 16 4 9 12 16 5 6 12 16 5 7 12 16 5 8 12 16 5 9 12 16 1 6 13 16 17 13 16 1 8 13 16 1 9 13 16 2 6 13 16 2 7 13 16 2 8 13 16 2 9 13 16 3 613 16 3 7 13 16 3 8 13 16 3 9 13 16 4 6 13 16 4 7 13 16 4 8 13 16 4 9 1316 5 6 13 16 5 7 13 16 5 8 13 16 5 9 13 16 1 6 10 17 1 7 10 17 1 8 10 171 9 10 17 2 6 10 17 2 7 10 17 2 8 10 17 2 9 10 17 3 6 10 17 3 7 10 17 38 10 17 3 9 10 17 4 6 10 17 4 7 10 17 4 8 10 17 4 9 10 17 5 6 10 17 5 710 17 5 8 10 17 5 9 10 17 1 6 11 17 1 7 11 17 1 8 11 17 1 9 11 17 2 6 1117 2 7 11 17 2 8 11 17 2 9 11 17 3 6 11 17 3 7 11 17 3 8 11 17 3 9 11 174 6 11 17 4 7 11 17 4 8 11 17 4 9 11 17 5 6 11 17 5 7 11 17 5 8 11 17 59 11 17 1 6 12 17 1 7 12 17 1 8 12 17 1 9 12 17 2 6 12 17 2 7 12 17 2 812 17 2 9 12 17 3 6 12 17 3 7 12 17 3 8 12 17 3 9 12 17 4 6 12 17 4 7 1217 4 8 12 17 4 9 12 17 5 6 12 17 5 7 12 17 5 8 12 17 5 9 12 17 1 6 13 171 7 13 17 1 8 13 17 1 9 13 17 2 6 13 17 2 7 13 17 2 8 13 17 2 9 13 17 36 13 17 3 7 13 17 3 8 13 17 3 9 13 17 4 6 13 17 4 7 13 17 4 8 13 17 4 913 17 5 6 13 17 5 7 13 17 5 8 13 17 5 9 13 17 1 6 10 18 1 7 10 18 1 8 1018 1 9 10 18 2 6 10 18 2 7 10 18 2 8 10 18 2 9 10 18 3 6 10 18 3 7 10 183 8 10 18 3 9 10 18 4 6 10 18 4 7 10 18 4 8 10 18 4 9 10 18 5 6 10 18 57 10 18 5 8 10 18 5 9 10 18 1 6 11 18 1 7 11 18 1 8 11 18 1 9 11 18 2 611 18 2 7 11 18 2 8 11 18 2 9 11 18 3 6 11 18 3 7 11 18 3 8 11 18 3 9 1118 4 6 11 18 4 7 11 18 4 8 11 18 4 9 11 18 5 6 11 18 5 7 11 18 5 8 11 185 9 11 18 1 6 12 18 1 7 12 18 1 8 12 18 1 9 12 18 2 6 12 18 2 7 12 18 28 12 18 2 9 12 18 3 6 12 18 3 7 12 18 3 8 12 18 3 9 12 18 4 6 12 18 4 712 18 4 8 12 18 4 9 12 18 5 6 12 18 5 7 12 18 5 8 12 18 5 9 12 18 1 6 1318 1 7 13 18 1 8 13 18 1 9 13 18 2 6 13 18 2 7 13 18 2 8 13 18 2 9 13 183 6 13 18 3 7 13 18 3 8 13 18 3 9 13 18 4 6 13 18 4 7 13 18 4 8 13 18 49 13 18 5 6 13 18 5 7 13 18 5 8 13 18 5 9 13 18 1 6 10 19 1 7 10 19 1 810 19 1 9 10 19 2 6 10 19 2 7 10 19 2 8 10 19 2 9 10 19 3 6 10 19 3 7 1019 3 8 10 19 3 9 10 19 4 6 10 19 4 7 10 19 4 8 10 19 4 9 10 19 5 6 10 195 7 10 19 5 8 10 19 5 9 10 19 1 6 11 19 1 7 11 19 1 8 11 19 1 9 11 19 26 11 19 2 7 11 19 2 8 11 19 2 9 11 19 3 6 11 19 3 7 11 19 3 8 11 19 3 911 19 4 6 11 19 4 7 11 19 4 8 11 19 4 9 11 19 5 6 11 19 5 7 11 19 5 8 1119 5 9 11 19 1 6 12 19 1 7 12 19 1 8 12 19 1 9 12 19 2 6 12 19 2 7 12 192 8 12 19 2 9 12 19 3 6 12 19 3 7 12 19 3 8 12 19 3 9 12 19 4 6 12 19 47 12 19 4 8 12 19 4 9 12 19 5 6 12 19 5 7 12 19 5 8 12 19 5 9 12 19 1 613 19 1 7 13 19 1 8 13 19 1 9 13 19 2 6 13 19 2 7 13 19 2 8 13 19 2 9 1319 3 6 13 19 3 7 13 19 3 8 13 19 3 9 13 19 4 6 13 19 4 7 13 19 4 8 13 194 9 13 19 5 6 13 19 5 7 13 19 5 8 13 19 5 9 13 19 1 6 10 20 1 7 10 20 18 10 20 1 9 10 20 2 6 10 20 2 7 10 20 2 8 10 20 2 9 10 20 3 6 10 20 3 710 20 3 8 10 20 3 9 10 20 4 6 10 20 4 7 10 20 4 8 10 20 4 9 10 20 5 6 1020 5 7 10 20 5 8 10 20 5 9 10 20 1 6 11 20 1 7 11 20 1 8 11 20 1 9 11 202 6 11 20 2 7 11 20 2 8 11 20 2 9 11 20 3 6 11 20 3 7 11 20 3 8 11 20 39 11 20 4 6 11 20 4 7 11 20 4 8 11 20 4 9 11 20 5 6 11 20 5 7 11 20 5 811 20 5 9 11 20 1 6 12 20 1 7 12 20 1 8 12 20 1 9 12 20 2 6 12 20 2 7 1220 2 8 12 20 2 9 12 20 3 6 12 20 3 7 12 20 3 8 12 20 3 9 12 20 4 6 12 204 7 12 20 4 8 12 20 4 9 12 20 5 6 12 20 5 7 12 20 5 8 12 20 5 9 12 20 16 13 20 1 7 13 20 1 8 13 20 1 9 13 20 2 6 13 20 2 7 13 20 2 8 13 20 2 913 20 3 6 13 20 3 7 13 20 3 8 13 20 3 9 13 20 4 6 13 20 4 7 13 20 4 8 1320 4 9 13 20 5 6 13 20 5 7 13 20 5 8 13 20 5 9 13 20 1 6 10 21 1 7 10 211 8 10 21 1 9 10 21 2 6 10 21 2 7 10 21 2 8 10 21 2 9 10 21 3 6 10 21 37 10 21 3 8 10 21 3 9 10 21 4 6 10 21 4 7 10 21 4 8 10 21 4 9 10 21 5 610 21 5 7 10 21 5 8 10 21 5 9 10 21 1 6 11 21 1 7 11 21 1 8 11 21 1 9 1121 2 6 11 21 2 7 11 21 2 8 11 21 2 9 11 21 3 6 11 21 3 7 11 21 3 8 11 213 9 11 21 4 6 11 21 4 7 11 21 4 8 11 21 4 9 11 21 5 6 11 21 5 7 11 21 58 11 21 5 9 11 21 1 6 12 21 1 7 12 21 1 8 12 21 1 9 12 21 2 6 12 21 2 712 21 2 8 12 21 2 9 12 21 3 6 12 21 3 7 12 21 3 8 12 21 3 9 12 21 4 6 1221 4 7 12 21 4 8 12 21 4 9 12 21 5 6 12 21 5 7 12 21 5 8 12 21 5 9 12 211 6 13 21 1 7 13 21 1 8 13 21 1 9 13 21 2 6 13 21 2 7 13 21 2 8 13 21 29 13 21 3 6 13 21 3 7 13 21 3 8 13 21 3 9 13 21 4 6 13 21 4 7 13 21 4 813 21 4 9 13 21 5 6 13 21 5 7 13 21 5 8 13 21 5 9 13 21 1 6 14 18 1 7 1418 1 8 14 18 1 9 14 18 2 6 14 18 2 7 14 18 2 8 14 18 2 9 14 18 3 6 14 183 7 14 18 3 8 14 18 3 9 14 18 4 6 14 18 4 7 14 18 4 8 14 18 4 9 14 18 56 14 18 5 7 14 18 5 8 14 18 5 9 14 18 1 6 15 18 1 7 15 18 1 8 15 18 1 915 18 2 6 15 18 2 7 15 18 2 8 15 18 2 9 15 18 3 6 15 18 3 7 15 18 3 8 1518 3 9 15 18 4 6 15 18 4 7 15 18 4 8 15 18 4 9 15 18 5 6 15 18 5 7 15 185 8 15 18 5 9 15 18 1 6 16 18 1 7 16 18 1 8 16 18 1 9 16 18 2 6 16 18 27 16 18 2 8 16 18 2 9 16 18 3 6 16 18 3 7 16 18 3 8 16 18 3 9 16 18 4 616 18 4 7 16 18 4 8 16 18 4 9 16 18 5 6 16 18 5 7 16 18 5 8 16 18 5 9 1618 1 6 17 18 1 7 17 18 1 8 17 18 1 9 17 18 2 6 17 18 2 7 17 18 2 8 17 182 9 17 18 3 6 17 18 3 7 17 18 3 8 17 18 3 9 17 18 4 6 17 18 4 7 17 18 48 17 18 4 9 17 18 5 6 17 18 5 7 17 18 5 8 17 18 5 9 17 18 1 6 14 19 1 714 19 1 8 14 19 1 9 14 19 2 6 14 19 2 7 14 19 2 8 14 19 2 9 14 19 3 6 1419 3 7 14 19 3 8 14 19 3 9 14 19 4 6 14 19 4 7 14 19 4 8 14 19 4 9 14 195 6 14 19 5 7 14 19 5 8 14 19 5 9 14 19 1 6 15 19 1 7 15 19 1 8 15 19 19 15 19 2 6 15 19 2 7 15 19 2 8 15 19 2 9 15 19 3 6 15 19 3 7 15 19 3 815 19 3 9 15 19 4 6 15 19 4 7 15 19 4 8 15 19 4 9 15 19 5 6 15 19 5 7 1519 5 8 15 19 5 9 15 19 1 6 16 19 1 7 16 19 1 8 16 19 1 9 16 19 2 6 16 192 7 16 19 2 8 16 19 2 9 16 19 3 6 16 19 3 7 16 19 3 8 16 19 3 9 16 19 46 16 19 4 7 16 19 4 8 16 19 4 9 16 19 5 6 16 19 5 7 16 19 5 8 16 19 5 916 19 1 6 17 19 1 7 17 19 1 8 17 19 1 9 17 19 2 6 17 19 2 7 17 19 2 8 1719 2 9 17 19 3 6 17 19 3 7 17 19 3 8 17 19 3 9 17 19 4 6 17 19 4 7 17 194 8 17 19 4 9 17 19 5 6 17 19 5 7 17 19 5 8 17 19 5 9 17 19 1 6 14 20 17 14 20 1 8 14 20 1 9 14 20 2 6 14 20 2 7 14 20 2 8 14 20 2 9 14 20 3 614 20 3 7 14 20 3 8 14 20 3 9 14 20 4 6 14 20 4 7 14 20 4 8 14 20 4 9 1420 5 6 14 20 5 7 14 20 5 8 14 20 5 9 14 20 1 6 15 20 1 7 15 20 1 8 15 201 9 15 20 2 6 15 20 2 7 15 20 2 8 15 20 2 9 15 20 3 6 15 20 3 7 15 20 38 15 20 3 9 15 20 4 6 15 20 4 7 15 20 4 8 15 20 4 9 15 20 5 6 15 20 5 715 20 5 8 15 20 5 9 15 20 1 6 16 20 1 7 16 20 1 8 16 20 1 9 16 20 2 6 1620 2 7 16 20 2 8 16 20 2 9 16 20 3 6 16 20 3 7 16 20 3 8 16 20 3 9 16 204 6 16 20 4 7 16 20 4 8 16 20 4 9 16 20 5 6 16 20 5 7 16 20 5 8 16 20 59 16 20 1 6 17 20 1 7 17 20 1 8 17 20 1 9 17 20 2 6 17 20 2 7 17 20 2 817 20 2 9 17 20 3 6 17 20 3 7 17 20 3 8 17 20 3 9 17 20 4 6 17 20 4 7 1720 4 8 17 20 4 9 17 20 5 6 17 20 5 7 17 20 5 8 17 20 5 9 17 20 1 6 14 211 7 14 21 1 8 14 21 1 9 14 21 2 6 14 21 2 7 14 21 2 8 14 21 2 9 14 21 36 14 21 3 7 14 21 3 8 14 21 3 9 14 21 4 6 14 21 4 7 14 21 4 8 14 21 4 914 21 5 6 14 21 5 7 14 21 5 8 14 21 5 9 14 21 1 6 15 21 1 7 15 21 1 8 1521 1 9 15 21 2 6 15 21 2 7 15 21 2 8 15 21 2 9 15 21 3 6 15 21 3 7 15 213 8 15 21 3 9 15 21 4 6 15 21 4 7 15 21 4 8 15 21 4 9 15 21 5 6 15 21 57 15 21 5 8 15 21 5 9 15 21 1 6 16 21 1 7 16 21 1 8 16 21 1 9 16 21 2 616 21 2 7 16 21 2 8 16 21 2 9 16 21 3 6 16 21 3 7 16 21 3 8 16 21 3 9 1621 4 6 16 21 4 7 16 21 4 8 16 21 4 9 16 21 5 6 16 21 5 7 16 21 5 8 16 215 9 16 21 1 6 17 21 1 7 17 21 1 8 17 21 1 9 17 21 2 6 17 21 2 7 17 21 28 17 21 2 9 17 21 3 6 17 21 3 7 17 21 3 8 17 21 3 9 17 21 4 6 17 21 4 717 21 4 8 17 21 4 9 17 21 5 6 17 21 5 7 17 21 5 8 17 21 5 9 17 21 1 1014 18 1 11 14 18 1 12 14 18 1 13 14 18 2 10 14 18 2 11 14 18 2 12 14 182 13 14 18 3 10 14 18 3 11 14 18 3 12 14 18 3 13 14 18 4 10 14 18 4 1114 18 4 12 14 18 4 13 14 18 5 10 14 18 5 11 14 18 5 12 14 18 5 13 14 181 10 15 18 1 11 15 18 1 12 15 18 1 13 15 18 2 10 15 18 2 11 15 18 2 1215 18 2 13 15 18 3 10 15 18 3 11 15 18 3 12 15 18 3 13 15 18 4 10 15 184 11 15 18 4 12 15 18 4 13 15 18 5 10 15 18 5 11 15 18 5 12 15 18 5 1315 18 1 10 16 18 1 11 16 18 1 12 16 18 1 13 16 18 2 10 16 18 2 11 16 182 12 16 18 2 13 16 18 3 10 16 18 3 11 16 18 3 12 16 18 3 13 16 18 4 1016 18 4 11 16 18 4 12 16 18 4 13 16 18 5 10 16 18 5 11 16 18 5 12 16 185 13 16 18 1 10 17 18 1 11 17 18 1 12 17 18 1 13 17 18 2 10 17 18 2 1117 18 2 12 17 18 2 13 17 18 3 10 17 18 3 11 17 18 3 12 17 18 3 13 17 184 10 17 18 4 11 17 18 4 12 17 18 4 13 17 18 5 10 17 18 5 11 17 18 5 1217 18 5 13 17 18 1 10 14 19 1 11 14 19 1 12 14 19 1 13 14 19 2 10 14 192 11 14 19 2 12 14 19 2 13 14 19 3 10 14 19 3 11 14 19 3 12 14 19 3 1314 19 4 10 14 19 4 11 14 19 4 12 14 19 4 13 14 19 5 10 14 19 5 11 14 195 12 14 19 5 13 14 19 1 10 15 19 1 11 15 19 1 12 15 19 1 13 15 19 2 1015 19 2 11 15 19 2 12 15 19 2 13 15 19 3 10 15 19 3 11 15 19 3 12 15 193 13 15 19 4 10 15 19 4 11 15 19 4 12 15 19 4 13 15 19 5 10 15 19 5 1115 19 5 12 15 19 5 13 15 19 1 10 16 19 1 11 16 19 1 12 16 19 1 13 16 192 10 16 19 2 11 16 19 2 12 16 19 2 13 16 19 3 10 16 19 3 11 16 19 3 1216 19 3 13 16 19 4 10 16 19 4 11 16 19 4 12 16 19 4 13 16 19 5 10 16 195 11 16 19 5 12 16 19 5 13 16 19 1 10 17 19 1 11 17 19 1 12 17 19 1 1317 19 2 10 17 19 2 11 17 19 2 12 17 19 2 13 17 19 3 10 17 19 3 11 17 193 12 17 19 3 13 17 19 4 10 17 19 4 11 17 19 4 12 17 19 4 13 17 19 5 1017 19 5 11 17 19 5 12 17 19 5 13 17 19 1 10 14 20 1 11 14 20 1 12 14 201 13 14 20 2 10 14 20 2 11 14 20 2 12 14 20 2 13 14 20 3 10 14 20 3 1114 20 3 12 14 20 3 13 14 20 4 10 14 20 4 11 14 20 4 12 14 20 4 13 14 205 10 14 20 5 11 14 20 5 12 14 20 5 13 14 20 1 10 15 20 1 11 15 20 1 1215 20 1 13 15 20 2 10 15 20 2 11 15 20 2 12 15 20 2 13 15 20 3 10 15 203 11 15 20 3 12 15 20 3 13 15 20 4 10 15 20 4 11 15 20 4 12 15 20 4 1315 20 5 10 15 20 5 11 15 20 5 12 15 20 5 13 15 20 1 10 16 20 1 11 16 201 12 16 20 1 13 16 20 2 10 16 20 2 11 16 20 2 12 16 20 2 13 16 20 3 1016 20 3 11 16 20 3 12 16 20 3 13 16 20 4 10 16 20 4 11 16 20 4 12 16 204 13 16 20 5 10 16 20 5 11 16 20 5 12 16 20 5 13 16 20 1 10 17 20 1 1117 20 1 12 17 20 1 13 17 20 2 10 17 20 2 11 17 20 2 12 17 20 2 13 17 203 10 17 20 3 11 17 20 3 12 17 20 3 13 17 20 4 10 17 20 4 11 17 20 4 1217 20 4 13 17 20 5 10 17 20 5 11 17 20 5 12 17 20 5 13 17 20 1 10 14 211 11 14 21 1 12 14 21 1 13 14 21 2 10 14 21 2 11 14 21 2 12 14 21 2 1314 21 3 10 14 21 3 11 14 21 3 12 14 21 3 13 14 21 4 10 14 21 4 11 14 214 12 14 21 4 13 14 21 5 10 14 21 5 11 14 21 5 12 14 21 5 13 14 21 1 1015 21 1 11 15 21 1 12 15 21 1 13 15 21 2 10 15 21 2 11 15 21 2 12 15 212 13 15 21 3 10 15 21 3 11 15 21 3 12 15 21 3 13 15 21 4 10 15 21 4 1115 21 4 12 15 21 4 13 15 21 5 10 15 21 5 11 15 21 5 12 15 21 5 13 15 211 10 16 21 1 11 16 21 1 12 16 21 1 13 16 21 2 10 16 21 2 11 16 21 2 1216 21 2 13 16 21 3 10 16 21 3 11 16 21 3 12 16 21 3 13 16 21 4 10 16 214 11 16 21 4 12 16 21 4 13 16 21 5 10 16 21 5 11 16 21 5 12 16 21 5 1316 21 1 10 17 21 1 11 17 21 1 12 17 21 1 13 17 21 2 10 17 21 2 11 17 212 12 17 21 2 13 17 21 3 10 17 21 3 11 17 21 3 12 17 21 3 13 17 21 4 1017 21 4 11 17 21 4 12 17 21 4 13 17 21 5 10 17 21 5 11 17 21 5 12 17 215 13 17 21 6 10 14 18 6 11 14 18 6 12 14 18 6 13 14 18 7 10 14 18 7 1114 18 7 12 14 18 7 13 14 18 8 10 14 18 8 11 14 18 8 12 14 18 8 13 14 189 10 14 18 9 11 14 18 9 12 14 18 9 13 14 18 6 10 15 18 6 11 15 18 6 1215 18 6 13 15 18 7 10 15 18 7 11 15 18 7 12 15 18 7 13 15 18 8 10 15 188 11 15 18 8 12 15 18 8 13 15 18 9 10 15 18 9 11 15 18 9 12 15 18 9 1315 18 6 10 16 18 6 11 16 18 6 12 16 18 6 13 16 18 7 10 16 18 7 11 16 187 12 16 18 7 13 16 18 8 10 16 18 8 11 16 18 8 12 16 18 8 13 16 18 9 1016 18 9 11 16 18 9 12 16 18 9 13 16 18 6 10 17 18 6 11 17 18 6 12 17 186 13 17 18 7 10 17 18 7 11 17 18 7 12 17 18 7 13 17 18 8 10 17 18 8 1117 18 8 12 17 18 8 13 17 18 9 10 17 18 9 11 17 18 9 12 17 18 9 13 17 186 10 14 19 6 11 14 19 6 12 14 19 6 13 14 19 7 10 14 19 7 11 14 19 7 1214 19 7 13 14 19 8 10 14 19 8 11 14 19 8 12 14 19 8 13 14 19 9 10 14 199 11 14 19 9 12 14 19 9 13 14 19 6 10 15 19 6 11 15 19 6 12 15 19 6 1315 19 7 10 15 19 7 11 15 19 7 12 15 19 7 13 15 19 8 10 15 19 8 11 15 198 12 15 19 8 13 15 19 9 10 15 19 9 11 15 19 9 12 15 19 9 13 15 19 6 1016 19 6 11 16 19 6 12 16 19 6 13 16 19 7 10 16 19 7 11 16 19 7 12 16 197 13 16 19 8 10 16 19 8 11 16 19 8 12 16 19 8 13 16 19 9 10 16 19 9 1116 19 9 12 16 19 9 13 16 19 6 10 17 19 6 11 17 19 6 12 17 19 6 13 17 197 10 17 19 7 11 17 19 7 12 17 19 7 13 17 19 8 10 17 19 8 11 17 19 8 1217 19 8 13 17 19 9 10 17 19 9 11 17 19 9 12 17 19 9 13 17 19 6 10 14 206 11 14 20 6 12 14 20 6 13 14 20 7 10 14 20 7 11 14 20 7 12 14 20 7 1314 20 8 10 14 20 8 11 14 20 8 12 14 20 8 13 14 20 9 10 14 20 9 11 14 209 12 14 20 9 13 14 20 6 10 15 20 6 11 15 20 6 12 15 20 6 13 15 20 7 1015 20 7 11 15 20 7 12 15 20 7 13 15 20 8 10 15 20 8 11 15 20 8 12 15 208 13 15 20 9 10 15 20 9 11 15 20 9 12 15 20 9 13 15 20 6 10 16 20 6 1116 20 6 12 16 20 6 13 16 20 7 10 16 20 7 11 16 20 7 12 16 20 7 13 16 208 10 16 20 8 11 16 20 8 12 16 20 8 13 16 20 9 10 16 20 9 11 16 20 9 1216 20 9 13 16 20 6 10 17 20 6 11 17 20 6 12 17 20 6 13 17 20 7 10 17 207 11 17 20 7 12 17 20 7 13 17 20 8 10 17 20 8 11 17 20 8 12 17 20 8 1317 20 9 10 17 20 9 11 17 20 9 12 17 20 9 13 17 20 6 10 14 21 6 11 14 216 12 14 21 6 13 14 21 7 10 14 21 7 11 14 21 7 12 14 21 7 13 14 21 8 1014 21 8 11 14 21 8 12 14 21 8 13 14 21 9 10 14 21 9 11 14 21 9 12 14 219 13 14 21 6 10 15 21 6 11 15 21 6 12 15 21 6 13 15 21 7 10 15 21 7 1115 21 7 12 15 21 7 13 15 21 8 10 15 21 8 11 15 21 8 12 15 21 8 13 15 219 10 15 21 9 11 15 21 9 12 15 21 9 13 15 21 6 10 16 21 6 11 16 21 6 1216 21 6 13 16 21 7 10 16 21 7 11 16 21 7 12 16 21 7 13 16 21 8 10 16 218 11 16 21 8 12 16 21 8 13 16 21 9 10 16 21 9 11 16 21 9 12 16 21 9 1316 21 6 10 17 21 6 11 17 21 6 12 17 21 6 13 17 21 7 10 17 21 7 11 17 217 12 17 21 7 13 17 21 8 10 17 21 8 11 17 21 8 12 17 21 8 13 17 21 9 1017 21 9 11 17 21 9 12 17 21 9 13 17 21

TABLE 6 Five-way combinations of hematopoietic stem cell populationparameters for assessment First Para- Second Para- Third Para- FourthPara- Fifth Para- meter No. meter No. meter No. meter No. meter No. 1 610 14 18 1 7 10 14 18 1 8 10 14 18 1 9 10 14 18 2 6 10 14 18 2 7 10 1418 2 8 10 14 18 2 9 10 14 18 3 6 10 14 18 3 7 10 14 18 3 8 10 14 18 3 910 14 18 4 6 10 14 18 4 7 10 14 18 4 8 10 14 18 4 9 10 14 18 5 6 10 1418 5 7 10 14 18 5 8 10 14 18 5 9 10 14 18 1 6 11 14 18 1 7 11 14 18 1 811 14 18 1 9 11 14 18 2 6 11 14 18 2 7 11 14 18 2 8 11 14 18 2 9 11 1418 3 6 11 14 18 3 7 11 14 18 3 8 11 14 18 3 9 11 14 18 4 6 11 14 18 4 711 14 18 4 8 11 14 18 4 9 11 14 18 5 6 11 14 18 5 7 11 14 18 5 8 11 1418 5 9 11 14 18 1 6 12 14 18 1 7 12 14 18 1 8 12 14 18 1 9 12 14 18 2 612 14 18 2 7 12 14 18 2 8 12 14 18 2 9 12 14 18 3 6 12 14 18 3 7 12 1418 3 8 12 14 18 3 9 12 14 18 4 6 12 14 18 4 7 12 14 18 4 8 12 14 18 4 912 14 18 5 6 12 14 18 5 7 12 14 18 5 8 12 14 18 5 9 12 14 18 1 6 13 1418 1 7 13 14 18 1 8 13 14 18 1 9 13 14 18 2 6 13 14 18 2 7 13 14 18 2 813 14 18 2 9 13 14 18 3 6 13 14 18 3 7 13 14 18 3 8 13 14 18 3 9 13 1418 4 6 13 14 18 4 7 13 14 18 4 8 13 14 18 4 9 13 14 18 5 6 13 14 18 5 713 14 18 5 8 13 14 18 5 9 13 14 18 1 6 10 15 18 1 7 10 15 18 1 8 10 1518 1 9 10 15 18 2 6 10 15 18 2 7 10 15 18 2 8 10 15 18 2 9 10 15 18 3 610 15 18 3 7 10 15 18 3 8 10 15 18 3 9 10 15 18 4 6 10 15 18 4 7 10 1518 4 8 10 15 18 4 9 10 15 18 5 6 10 15 18 5 7 10 15 18 5 8 10 15 18 5 910 15 18 1 6 11 15 18 1 7 11 15 18 1 8 11 15 18 1 9 11 15 18 2 6 11 1518 2 7 11 15 18 2 8 11 15 18 2 9 11 15 18 3 6 11 15 18 3 7 11 15 18 3 811 15 18 3 9 11 15 18 4 6 11 15 18 4 7 11 15 18 4 8 11 15 18 4 9 11 1518 5 6 11 15 18 5 7 11 15 18 5 8 11 15 18 5 9 11 15 18 1 6 12 15 18 1 712 15 18 1 8 12 15 18 1 9 12 15 18 2 6 12 15 18 2 7 12 15 18 2 8 12 1518 2 9 12 15 18 3 6 12 15 18 3 7 12 15 18 3 8 12 15 18 3 9 12 15 18 4 612 15 18 4 7 12 15 18 4 8 12 15 18 4 9 12 15 18 5 6 12 15 18 5 7 12 1518 5 8 12 15 18 5 9 12 15 18 1 6 13 15 18 1 7 13 15 18 1 8 13 15 18 1 913 15 18 2 6 13 15 18 2 7 13 15 18 2 8 13 15 18 2 9 13 15 18 3 6 13 1518 3 7 13 15 18 3 8 13 15 18 3 9 13 15 18 4 6 13 15 18 4 7 13 15 18 4 813 15 18 4 9 13 15 18 5 6 13 15 18 5 7 13 15 18 5 8 13 15 18 5 9 13 1518 1 6 10 16 18 1 7 10 16 18 1 8 10 16 18 1 9 10 16 18 2 6 10 16 18 2 710 16 18 2 8 10 16 18 2 9 10 16 18 3 6 10 16 18 3 7 10 16 18 3 8 10 1618 3 9 10 16 18 4 6 10 16 18 4 7 10 16 18 4 8 10 16 18 4 9 10 16 18 5 610 16 18 5 7 10 16 18 5 8 10 16 18 5 9 10 16 18 1 6 11 16 18 1 7 11 1618 1 8 11 16 18 1 9 11 16 18 2 6 11 16 18 2 7 11 16 18 2 8 11 16 18 2 911 16 18 3 6 11 16 18 3 7 11 16 18 3 8 11 16 18 3 9 11 16 18 4 6 11 1618 4 7 11 16 18 4 8 11 16 18 4 9 11 16 18 5 6 11 16 18 5 7 11 16 18 5 811 16 18 5 9 11 16 18 1 6 12 16 18 1 7 12 16 18 1 8 12 16 18 1 9 12 1618 2 6 12 16 18 2 7 12 16 18 2 8 12 16 18 2 9 12 16 18 3 6 12 16 18 3 712 16 18 3 8 12 16 18 3 9 12 16 18 4 6 12 16 18 4 7 12 16 18 4 8 12 1618 4 9 12 16 18 5 6 12 16 18 5 7 12 16 18 5 8 12 16 18 5 9 12 16 18 1 613 16 18 1 7 13 16 18 1 8 13 16 18 1 9 13 16 18 2 6 13 16 18 2 7 13 1618 2 8 13 16 18 2 9 13 16 18 3 6 13 16 18 3 7 13 16 18 3 8 13 16 18 3 913 16 18 4 6 13 16 18 4 7 13 16 18 4 8 13 16 18 4 9 13 16 18 5 6 13 1618 5 7 13 16 18 5 8 13 16 18 5 9 13 16 18 1 6 10 17 18 1 7 10 17 18 1 810 17 18 1 9 10 17 18 2 6 10 17 18 2 7 10 17 18 2 8 10 17 18 2 9 10 1718 3 6 10 17 18 3 7 10 17 18 3 8 10 17 18 3 9 10 17 18 4 6 10 17 18 4 710 17 18 4 8 10 17 18 4 9 10 17 18 5 6 10 17 18 5 7 10 17 18 5 8 10 1718 5 9 10 17 18 1 6 11 17 18 1 7 11 17 18 1 8 11 17 18 1 9 11 17 18 2 611 17 18 2 7 11 17 18 2 8 11 17 18 2 9 11 17 18 3 6 11 17 18 3 7 11 1718 3 8 11 17 18 3 9 11 17 18 4 6 11 17 18 4 7 11 17 18 4 8 11 17 18 4 911 17 18 5 6 11 17 18 5 7 11 17 18 5 8 11 17 18 5 9 11 17 18 1 6 12 1718 1 7 12 17 18 1 8 12 17 18 1 9 12 17 18 2 6 12 17 18 2 7 12 17 18 2 812 17 18 2 9 12 17 18 3 6 12 17 18 3 7 12 17 18 3 8 12 17 18 3 9 12 1718 4 6 12 17 18 4 7 12 17 18 4 8 12 17 18 4 9 12 17 18 5 6 12 17 18 5 712 17 18 5 8 12 17 18 5 9 12 17 18 1 6 13 17 18 1 7 13 17 18 1 8 13 1718 1 9 13 17 18 2 6 13 17 18 2 7 13 17 18 2 8 13 17 18 2 9 13 17 18 3 613 17 18 3 7 13 17 18 3 8 13 17 18 3 9 13 17 18 4 6 13 17 18 4 7 13 1718 4 8 13 17 18 4 9 13 17 18 5 6 13 17 18 5 7 13 17 18 5 8 13 17 18 5 913 17 18 1 6 10 14 19 1 7 10 14 19 1 8 10 14 19 1 9 10 14 19 2 6 10 1419 2 7 10 14 19 2 8 10 14 19 2 9 10 14 19 3 6 10 14 19 3 7 10 14 19 3 810 14 19 3 9 10 14 19 4 6 10 14 19 4 7 10 14 19 4 8 10 14 19 4 9 10 1419 5 6 10 14 19 5 7 10 14 19 5 8 10 14 19 5 9 10 14 19 1 6 11 14 19 1 711 14 19 1 8 11 14 19 1 9 11 14 19 2 6 11 14 19 2 7 11 14 19 2 8 11 1419 2 9 11 14 19 3 6 11 14 19 3 7 11 14 19 3 8 11 14 19 3 9 11 14 19 4 611 14 19 4 7 11 14 19 4 8 11 14 19 4 9 11 14 19 5 6 11 14 19 5 7 11 1419 5 8 11 14 19 5 9 11 14 19 1 6 12 14 19 1 7 12 14 19 1 8 12 14 19 1 912 14 19 2 6 12 14 19 2 7 12 14 19 2 8 12 14 19 2 9 12 14 19 3 6 12 1419 3 7 12 14 19 3 8 12 14 19 3 9 12 14 19 4 6 12 14 19 4 7 12 14 19 4 812 14 19 4 9 12 14 19 5 6 12 14 19 5 7 12 14 19 5 8 12 14 19 5 9 12 1419 1 6 13 14 19 1 7 13 14 19 1 8 13 14 19 1 9 13 14 19 2 6 13 14 19 2 713 14 19 2 8 13 14 19 2 9 13 14 19 3 6 13 14 19 3 7 13 14 19 3 8 13 1419 3 9 13 14 19 4 6 13 14 19 4 7 13 14 19 4 8 13 14 19 4 9 13 14 19 5 613 14 19 5 7 13 14 19 5 8 13 14 19 5 9 13 14 19 1 6 10 15 19 1 7 10 1519 1 8 10 15 19 1 9 10 15 19 2 6 10 15 19 2 7 10 15 19 2 8 10 15 19 2 910 15 19 3 6 10 15 19 3 7 10 15 19 3 8 10 15 19 3 9 10 15 19 4 6 10 1519 4 7 10 15 19 4 8 10 15 19 4 9 10 15 19 5 6 10 15 19 5 7 10 15 19 5 810 15 19 5 9 10 15 19 1 6 11 15 19 1 7 11 15 19 1 8 11 15 19 1 9 11 1519 2 6 11 15 19 2 7 11 15 19 2 8 11 15 19 2 9 11 15 19 3 6 11 15 19 3 711 15 19 3 8 11 15 19 3 9 11 15 19 4 6 11 15 19 4 7 11 15 19 4 8 11 1519 4 9 11 15 19 5 6 11 15 19 5 7 11 15 19 5 8 11 15 19 5 9 11 15 19 1 612 15 19 1 7 12 15 19 1 8 12 15 19 1 9 12 15 19 2 6 12 15 19 2 7 12 1519 2 8 12 15 19 2 9 12 15 19 3 6 12 15 19 3 7 12 15 19 3 8 12 15 19 3 912 15 19 4 6 12 15 19 4 7 12 15 19 4 8 12 15 19 4 9 12 15 19 5 6 12 1519 5 7 12 15 19 5 8 12 15 19 5 9 12 15 19 1 6 13 15 19 1 7 13 15 19 1 813 15 19 1 9 13 15 19 2 6 13 15 19 2 7 13 15 19 2 8 13 15 19 2 9 13 1519 3 6 13 15 19 3 7 13 15 19 3 8 13 15 19 3 9 13 15 19 4 6 13 15 19 4 713 15 19 4 8 13 15 19 4 9 13 15 19 5 6 13 15 19 5 7 13 15 19 5 8 13 1519 5 9 13 15 19 1 6 10 16 19 1 7 10 16 19 1 8 10 16 19 1 9 10 16 19 2 610 16 19 2 7 10 16 19 2 8 10 16 19 2 9 10 16 19 3 6 10 16 19 3 7 10 1619 3 8 10 16 19 3 9 10 16 19 4 6 10 16 19 4 7 10 16 19 4 8 10 16 19 4 910 16 19 5 6 10 16 19 5 7 10 16 19 5 8 10 16 19 5 9 10 16 19 1 6 11 1619 1 7 11 16 19 1 8 11 16 19 1 9 11 16 19 2 6 11 16 19 2 7 11 16 19 2 811 16 19 2 9 11 16 19 3 6 11 16 19 3 7 11 16 19 3 8 11 16 19 3 9 11 1619 4 6 11 16 19 4 7 11 16 19 4 8 11 16 19 4 9 11 16 19 5 6 11 16 19 5 711 16 19 5 8 11 16 19 5 9 11 16 19 1 6 12 16 19 1 7 12 16 19 1 8 12 1619 1 9 12 16 19 2 6 12 16 19 2 7 12 16 19 2 8 12 16 19 2 9 12 16 19 3 612 16 19 3 7 12 16 19 3 8 12 16 19 3 9 12 16 19 4 6 12 16 19 4 7 12 1619 4 8 12 16 19 4 9 12 16 19 5 6 12 16 19 5 7 12 16 19 5 8 12 16 19 5 912 16 19 1 6 13 16 19 1 7 13 16 19 1 8 13 16 19 1 9 13 16 19 2 6 13 1619 2 7 13 16 19 2 8 13 16 19 2 9 13 16 19 3 6 13 16 19 3 7 13 16 19 3 813 16 19 3 9 13 16 19 4 6 13 16 19 4 7 13 16 19 4 8 13 16 19 4 9 13 1619 5 6 13 16 19 5 7 13 16 19 5 8 13 16 19 5 9 13 16 19 1 6 10 17 19 1 710 17 19 1 8 10 17 19 1 9 10 17 19 2 6 10 17 19 2 7 10 17 19 2 8 10 1719 2 9 10 17 19 3 6 10 17 19 3 7 10 17 19 3 8 10 17 19 3 9 10 17 19 4 610 17 19 4 7 10 17 19 4 8 10 17 19 4 9 10 17 19 5 6 10 17 19 5 7 10 1719 5 8 10 17 19 5 9 10 17 19 1 6 11 17 19 1 7 11 17 19 1 8 11 17 19 1 911 17 19 2 6 11 17 19 2 7 11 17 19 2 8 11 17 19 2 9 11 17 19 3 6 11 1719 3 7 11 17 19 3 8 11 17 19 3 9 11 17 19 4 6 11 17 19 4 7 11 17 19 4 811 17 19 4 9 11 17 19 5 6 11 17 19 5 7 11 17 19 5 8 11 17 19 5 9 11 1719 1 6 12 17 19 1 7 12 17 19 1 8 12 17 19 1 9 12 17 19 2 6 12 17 19 2 712 17 19 2 8 12 17 19 2 9 12 17 19 3 6 12 17 19 3 7 12 17 19 3 8 12 1719 3 9 12 17 19 4 6 12 17 19 4 7 12 17 19 4 8 12 17 19 4 9 12 17 19 5 612 17 19 5 7 12 17 19 5 8 12 17 19 5 9 12 17 19 1 6 13 17 19 1 7 13 1719 1 8 13 17 19 1 9 13 17 19 2 6 13 17 19 2 7 13 17 19 2 8 13 17 19 2 913 17 19 3 6 13 17 19 3 7 13 17 19 3 8 13 17 19 3 9 13 17 19 4 6 13 1719 4 7 13 17 19 4 8 13 17 19 4 9 13 17 19 5 6 13 17 19 5 7 13 17 19 5 813 17 19 5 9 13 17 19 1 6 10 14 20 1 7 10 14 20 1 8 10 14 20 1 9 10 1420 2 6 10 14 20 2 7 10 14 20 2 8 10 14 20 2 9 10 14 20 3 6 10 14 20 3 710 14 20 3 8 10 14 20 3 9 10 14 20 4 6 10 14 20 4 7 10 14 20 4 8 10 1420 4 9 10 14 20 5 6 10 14 20 5 7 10 14 20 5 8 10 14 20 5 9 10 14 20 1 611 14 20 1 7 11 14 20 1 8 11 14 20 1 9 11 14 20 2 6 11 14 20 2 7 11 1420 2 8 11 14 20 2 9 11 14 20 3 6 11 14 20 3 7 11 14 20 3 8 11 14 20 3 911 14 20 4 6 11 14 20 4 7 11 14 20 4 8 11 14 20 4 9 11 14 20 5 6 11 1420 5 7 11 14 20 5 8 11 14 20 5 9 11 14 20 1 6 12 14 20 1 7 12 14 20 1 812 14 20 1 9 12 14 20 2 6 12 14 20 2 7 12 14 20 2 8 12 14 20 2 9 12 1420 3 6 12 14 20 3 7 12 14 20 3 8 12 14 20 3 9 12 14 20 4 6 12 14 20 4 712 14 20 4 8 12 14 20 4 9 12 14 20 5 6 12 14 20 5 7 12 14 20 5 8 12 1420 5 9 12 14 20 1 6 13 14 20 1 7 13 14 20 1 8 13 14 20 1 9 13 14 20 2 613 14 20 2 7 13 14 20 2 8 13 14 20 2 9 13 14 20 3 6 13 14 20 3 7 13 1420 3 8 13 14 20 3 9 13 14 20 4 6 13 14 20 4 7 13 14 20 4 8 13 14 20 4 913 14 20 5 6 13 14 20 5 7 13 14 20 5 8 13 14 20 5 9 13 14 20 1 6 10 1520 1 7 10 15 20 1 8 10 15 20 1 9 10 15 20 2 6 10 15 20 2 7 10 15 20 2 810 15 20 2 9 10 15 20 3 6 10 15 20 3 7 10 15 20 3 8 10 15 20 3 9 10 1520 4 6 10 15 20 4 7 10 15 20 4 8 10 15 20 4 9 10 15 20 5 6 10 15 20 5 710 15 20 5 8 10 15 20 5 9 10 15 20 1 6 11 15 20 1 7 11 15 20 1 8 11 1520 1 9 11 15 20 2 6 11 15 20 2 7 11 15 20 2 8 11 15 20 2 9 11 15 20 3 611 15 20 3 7 11 15 20 3 8 11 15 20 3 9 11 15 20 4 6 11 15 20 4 7 11 1520 4 8 11 15 20 4 9 11 15 20 5 6 11 15 20 5 7 11 15 20 5 8 11 15 20 5 911 15 20 1 6 12 15 20 1 7 12 15 20 1 8 12 15 20 1 9 12 15 20 2 6 12 1520 2 7 12 15 20 2 8 12 15 20 2 9 12 15 20 3 6 12 15 20 3 7 12 15 20 3 812 15 20 3 9 12 15 20 4 6 12 15 20 4 7 12 15 20 4 8 12 15 20 4 9 12 1520 5 6 12 15 20 5 7 12 15 20 5 8 12 15 20 5 9 12 15 20 1 6 13 15 20 1 713 15 20 1 8 13 15 20 1 9 13 15 20 2 6 13 15 20 2 7 13 15 20 2 8 13 1520 2 9 13 15 20 3 6 13 15 20 3 7 13 15 20 3 8 13 15 20 3 9 13 15 20 4 613 15 20 4 7 13 15 20 4 8 13 15 20 4 9 13 15 20 5 6 13 15 20 5 7 13 1520 5 8 13 15 20 5 9 13 15 20 1 6 10 16 20 1 7 10 16 20 1 8 10 16 20 1 910 16 20 2 6 10 16 20 2 7 10 16 20 2 8 10 16 20 2 9 10 16 20 3 6 10 1620 3 7 10 16 20 3 8 10 16 20 3 9 10 16 20 4 6 10 16 20 4 7 10 16 20 4 810 16 20 4 9 10 16 20 5 6 10 16 20 5 7 10 16 20 5 8 10 16 20 5 9 10 1620 1 6 11 16 20 1 7 11 16 20 1 8 11 16 20 1 9 11 16 20 2 6 11 16 20 2 711 16 20 2 8 11 16 20 2 9 11 16 20 3 6 11 16 20 3 7 11 16 20 3 8 11 1620 3 9 11 16 20 4 6 11 16 20 4 7 11 16 20 4 8 11 16 20 4 9 11 16 20 5 611 16 20 5 7 11 16 20 5 8 11 16 20 5 9 11 16 20 1 6 12 16 20 1 7 12 1620 1 8 12 16 20 1 9 12 16 20 2 6 12 16 20 2 7 12 16 20 2 8 12 16 20 2 912 16 20 3 6 12 16 20 3 7 12 16 20 3 8 12 16 20 3 9 12 16 20 4 6 12 1620 4 7 12 16 20 4 8 12 16 20 4 9 12 16 20 5 6 12 16 20 5 7 12 16 20 5 812 16 20 5 9 12 16 20 1 6 13 16 20 1 7 13 16 20 1 8 13 16 20 1 9 13 1620 2 6 13 16 20 2 7 13 16 20 2 8 13 16 20 2 9 13 16 20 3 6 13 16 20 3 713 16 20 3 8 13 16 20 3 9 13 16 20 4 6 13 16 20 4 7 13 16 20 4 8 13 1620 4 9 13 16 20 5 6 13 16 20 5 7 13 16 20 5 8 13 16 20 5 9 13 16 20 1 610 17 20 1 7 10 17 20 1 8 10 17 20 1 9 10 17 20 2 6 10 17 20 2 7 10 1720 2 8 10 17 20 2 9 10 17 20 3 6 10 17 20 3 7 10 17 20 3 8 10 17 20 3 910 17 20 4 6 10 17 20 4 7 10 17 20 4 8 10 17 20 4 9 10 17 20 5 6 10 1720 5 7 10 17 20 5 8 10 17 20 5 9 10 17 20 1 6 11 17 20 1 7 11 17 20 1 811 17 20 1 9 11 17 20 2 6 11 17 20 2 7 11 17 20 2 8 11 17 20 2 9 11 1720 3 6 11 17 20 3 7 11 17 20 3 8 11 17 20 3 9 11 17 20 4 6 11 17 20 4 711 17 20 4 8 11 17 20 4 9 11 17 20 5 6 11 17 20 5 7 11 17 20 5 8 11 1720 5 9 11 17 20 1 6 12 17 20 1 7 12 17 20 1 8 12 17 20 1 9 12 17 20 2 612 17 20 2 7 12 17 20 2 8 12 17 20 2 9 12 17 20 3 6 12 17 20 3 7 12 1720 3 8 12 17 20 3 9 12 17 20 4 6 12 17 20 4 7 12 17 20 4 8 12 17 20 4 912 17 20 5 6 12 17 20 5 7 12 17 20 5 8 12 17 20 5 9 12 17 20 1 6 13 1720 1 7 13 17 20 1 8 13 17 20 1 9 13 17 20 2 6 13 17 20 2 7 13 17 20 2 813 17 20 2 9 13 17 20 3 6 13 17 20 3 7 13 17 20 3 8 13 17 20 3 9 13 1720 4 6 13 17 20 4 7 13 17 20 4 8 13 17 20 4 9 13 17 20 5 6 13 17 20 5 713 17 20 5 8 13 17 20 5 9 13 17 20 1 6 10 14 21 1 7 10 14 21 1 8 10 1421 1 9 10 14 21 2 6 10 14 21 2 7 10 14 21 2 8 10 14 21 2 9 10 14 21 3 610 14 21 3 7 10 14 21 3 8 10 14 21 3 9 10 14 21 4 6 10 14 21 4 7 10 1421 4 8 10 14 21 4 9 10 14 21 5 6 10 14 21 5 7 10 14 21 5 8 10 14 21 5 910 14 21 1 6 11 14 21 1 7 11 14 21 1 8 11 14 21 1 9 11 14 21 2 6 11 1421 2 7 11 14 21 2 8 11 14 21 2 9 11 14 21 3 6 11 14 21 3 7 11 14 21 3 811 14 21 3 9 11 14 21 4 6 11 14 21 4 7 11 14 21 4 8 11 14 21 4 9 11 1421 5 6 11 14 21 5 7 11 14 21 5 8 11 14 21 5 9 11 14 21 1 6 12 14 21 1 712 14 21 1 8 12 14 21 1 9 12 14 21 2 6 12 14 21 2 7 12 14 21 2 8 12 1421 2 9 12 14 21 3 6 12 14 21 3 7 12 14 21 3 8 12 14 21 3 9 12 14 21 4 612 14 21 4 7 12 14 21 4 8 12 14 21 4 9 12 14 21 5 6 12 14 21 5 7 12 1421 5 8 12 14 21 5 9 12 14 21 1 6 13 14 21 1 7 13 14 21 1 8 13 14 21 1 913 14 21 2 6 13 14 21 2 7 13 14 21 2 8 13 14 21 2 9 13 14 21 3 6 13 1421 3 7 13 14 21 3 8 13 14 21 3 9 13 14 21 4 6 13 14 21 4 7 13 14 21 4 813 14 21 4 9 13 14 21 5 6 13 14 21 5 7 13 14 21 5 8 13 14 21 5 9 13 1421 1 6 10 15 21 1 7 10 15 21 1 8 10 15 21 1 9 10 15 21 2 6 10 15 21 2 710 15 21 2 8 10 15 21 2 9 10 15 21 3 6 10 15 21 3 7 10 15 21 3 8 10 1521 3 9 10 15 21 4 6 10 15 21 4 7 10 15 21 4 8 10 15 21 4 9 10 15 21 5 610 15 21 5 7 10 15 21 5 8 10 15 21 5 9 10 15 21 1 6 11 15 21 1 7 11 1521 1 8 11 15 21 1 9 11 15 21 2 6 11 15 21 2 7 11 15 21 2 8 11 15 21 2 911 15 21 3 6 11 15 21 3 7 11 15 21 3 8 11 15 21 3 9 11 15 21 4 6 11 1521 4 7 11 15 21 4 8 11 15 21 4 9 11 15 21 5 6 11 15 21 5 7 11 15 21 5 811 15 21 5 9 11 15 21 1 6 12 15 21 1 7 12 15 21 1 8 12 15 21 1 9 12 1521 2 6 12 15 21 2 7 12 15 21 2 8 12 15 21 2 9 12 15 21 3 6 12 15 21 3 712 15 21 3 8 12 15 21 3 9 12 15 21 4 6 12 15 21 4 7 12 15 21 4 8 12 1521 4 9 12 15 21 5 6 12 15 21 5 7 12 15 21 5 8 12 15 21 5 9 12 15 21 1 613 15 21 1 7 13 15 21 1 8 13 15 21 1 9 13 15 21 2 6 13 15 21 2 7 13 1521 2 8 13 15 21 2 9 13 15 21 3 6 13 15 21 3 7 13 15 21 3 8 13 15 21 3 913 15 21 4 6 13 15 21 4 7 13 15 21 4 8 13 15 21 4 9 13 15 21 5 6 13 1521 5 7 13 15 21 5 8 13 15 21 5 9 13 15 21 1 6 10 16 21 1 7 10 16 21 1 810 16 21 1 9 10 16 21 2 6 10 16 21 2 7 10 16 21 2 8 10 16 21 2 9 10 1621 3 6 10 16 21 3 7 10 16 21 3 8 10 16 21 3 9 10 16 21 4 6 10 16 21 4 710 16 21 4 8 10 16 21 4 9 10 16 21 5 6 10 16 21 5 7 10 16 21 5 8 10 1621 5 9 10 16 21 1 6 11 16 21 1 7 11 16 21 1 8 11 16 21 1 9 11 16 21 2 611 16 21 2 7 11 16 21 2 8 11 16 21 2 9 11 16 21 3 6 11 16 21 3 7 11 1621 3 8 11 16 21 3 9 11 16 21 4 6 11 16 21 4 7 11 16 21 4 8 11 16 21 4 911 16 21 5 6 11 16 21 5 7 11 16 21 5 8 11 16 21 5 9 11 16 21 1 6 12 1621 1 7 12 16 21 1 8 12 16 21 1 9 12 16 21 2 6 12 16 21 2 7 12 16 21 2 812 16 21 2 9 12 16 21 3 6 12 16 21 3 7 12 16 21 3 8 12 16 21 3 9 12 1621 4 6 12 16 21 4 7 12 16 21 4 8 12 16 21 4 9 12 16 21 5 6 12 16 21 5 712 16 21 5 8 12 16 21 5 9 12 16 21 1 6 13 16 21 1 7 13 16 21 1 8 13 1621 1 9 13 16 21 2 6 13 16 21 2 7 13 16 21 2 8 13 16 21 2 9 13 16 21 3 613 16 21 3 7 13 16 21 3 8 13 16 21 3 9 13 16 21 4 6 13 16 21 4 7 13 1621 4 8 13 16 21 4 9 13 16 21 5 6 13 16 21 5 7 13 16 21 5 8 13 16 21 5 913 16 21 1 6 10 17 21 1 7 10 17 21 1 8 10 17 21 1 9 10 17 21 2 6 10 1721 2 7 10 17 21 2 8 10 17 21 2 9 10 17 21 3 6 10 17 21 3 7 10 17 21 3 810 17 21 3 9 10 17 21 4 6 10 17 21 4 7 10 17 21 4 8 10 17 21 4 9 10 1721 5 6 10 17 21 5 7 10 17 21 5 8 10 17 21 5 9 10 17 21 1 6 11 17 21 1 711 17 21 1 8 11 17 21 1 9 11 17 21 2 6 11 17 21 2 7 11 17 21 2 8 11 1721 2 9 11 17 21 3 6 11 17 21 3 7 11 17 21 3 8 11 17 21 3 9 11 17 21 4 611 17 21 4 7 11 17 21 4 8 11 17 21 4 9 11 17 21 5 6 11 17 21 5 7 11 1721 5 8 11 17 21 5 9 11 17 21 1 6 12 17 21 1 7 12 17 21 1 8 12 17 21 1 912 17 21 2 6 12 17 21 2 7 12 17 21 2 8 12 17 21 2 9 12 17 21 3 6 12 1721 3 7 12 17 21 3 8 12 17 21 3 9 12 17 21 4 6 12 17 21 4 7 12 17 21 4 812 17 21 4 9 12 17 21 5 6 12 17 21 5 7 12 17 21 5 8 12 17 21 5 9 12 1721 1 6 13 17 21 1 7 13 17 21 1 8 13 17 21 1 9 13 17 21 2 6 13 17 21 2 713 17 21 2 8 13 17 21 2 9 13 17 21 3 6 13 17 21 3 7 13 17 21 3 8 13 1721 3 9 13 17 21 4 6 13 17 21 4 7 13 17 21 4 8 13 17 21 4 9 13 17 21 5 613 17 21 5 7 13 17 21 5 8 13 17 21 5 9 13 17 21

CXCR2 Agonists Gro-β, Gro-β T, and Variants Thereof

Exemplary CXCR2 agonists that may be used in conjunction with thecompositions and methods described herein are Gro-β and variantsthereof. Gro-β (also referred to as growth-regulated protein β,chemokine (C-X-C motif) ligand 2 (CXCL2), and macrophage inflammatoryprotein 2-α (MIP2-α)) is a cytokine capable of mobilizing hematopoieticstem and progenitor cells, for example, by stimulating the release ofproteases, and particularly MMP9, from peripheral neutrophils. Withoutbeing limited by mechanism, MMP9 may induce mobilization ofhematopoietic stem and progenitor cells from stem cell niches, such asthe bone marrow, to circulating peripheral blood by stimulating thedegradation of proteins such as stem cell factor, its correspondingreceptor, CD117, and CXCL12, all of which generally maintainhematopoietic stem and progenitor cells immobilized in bone marrow.

In addition to Gro-β, exemplary CXCR2 agonists that may be used inconjunction with the compositions and methods described herein aretruncated forms of Gro-β, such as those that feature a deletion at theN-terminus of Gro-β of from 1 to 8 amino acids (e.g., peptides thatfeature an N-terminal deletion of 1 amino acids, 2 amino acids, 3 aminoacids, 4 amino acids, 5 amino acids, 6 amino acids, 7 amino acids, or 8amino acids). In some embodiments, CXCR2 agonists that may be used inconjunction with the compositions and methods described herein includeGro-β T, which is characterized by a deletion of the first four aminoacids from the N-terminus of Gro-β. Gro-β T exhibits particularlyadvantageous biological properties, such as the ability to inducehematopoietic stem and progenitor cell mobilization with a potencysuperior to that of Gro-β by multiple orders of magnitude. Gro-β andGro-β T are described, for example, in U.S. Pat. No. 6,080,398, thedisclosure of which is incorporated herein by reference in its entirety.

In addition, exemplary CXCR2 agonists that may be used in conjunctionwith the compositions and methods described herein are variants of Gro-βcontaining an aspartic acid residue in place of the asparagine residueat position 69 of SEQ ID NO: 1. This peptide, referred to herein asGro-β N69D, retains the hematopoietic stem and progenitorcell-mobilizing functionality of Gro-β, yet induces this effect with asuperior potency. Similarly, CXCR2 agonists that may be used with thecompositions and methods described herein include variants of Gro-β Tcontaining an aspartic acid residue in place of the asparagine residueat position 65 of SEQ ID NO: 2. This peptide, referred to herein asGro-β T N65D, not only retains hematopoietic stem and progenitorcell-mobilizing capacity, but exhibits a potency that is substantiallygreater than that of Gro-β T. Gro-β N69D and Gro-β T N65D are described,for example, in U.S. Pat. No. 6,447,766, the disclosure of which isincorporated herein by reference in its entirety.

The amino acid sequences of Gro-β, Gro-β T, Gro-β N69D, and Gro-β T N65Dare set forth in TABLE 7, below.

TABLE 7 Amino acid sequences of Gro-β and select variants thereof SEQID NO. Description Amino Acid Sequence 1 Gro-β APLATELRCQCLQTLQGIHLKNIQSVKVKSPGPHCAQTEVIATL KNGQKACLNPASPMVKKIIEKM LKNGKSN 2 Gro-β-TTELRCQCLQTLQGIHLKNIQSV KVKSPGPHCAQTEVIATLKNGQ KACLNPASPMVKKIIEKMLKNG KSN3 Gro-β N69D APLATELRCQCLQTLQGIHLKN IQSVKVKSPGPHCAQTEVIATLKNGQKACLNPASPMVKKIIEKM LKDGKSN 4 Gro-β-T N65D TELRCQCLQTLQGIHLKNIQSVKVKSPGPHCAQTEVIATLKNGQ KACLNPASPMVKKIIEKMLKDG KSN

Additional CXCR2 agonists that may be used in conjunction with thecompositions and methods described herein include other variants ofGro-β, such as peptides that have one or more amino acid substitutions,insertions, and/or deletions relative to Gro-β. In some embodiments,CXCR2 agonists that may be used in conjunction with the compositions andmethods described herein include peptides having at least 85% sequenceidentity to the amino acid sequence of SEQ ID NO: 1 (e.g., a peptidehaving at least 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100%sequence identity to the amino acid sequence of SEQ ID NO: 1). In someembodiments, the amino acid sequence of the CXCR2 agonist differs fromthat of SEQ ID NO: 1 only by way of one or more conservative amino acidsubstitutions. In some embodiments, in some embodiments, the amino acidsequence of the CXCR2 agonist differs from that of SEQ ID NO: 1 by nomore than 20, no more than 15, no more than 10, no more than 5, or nomore than 1 nonconservative amino acid substitutions.

Additional examples of CXCR2 agonists useful in conjunction with thecompositions and methods described herein are variants of Gro-β T, suchas peptides that have one or more amino acid substitutions, insertions,and/or deletions relative to Gro-β T. In some embodiments, the CXCR2agonist may be a peptide having at least 85% sequence identity to theamino acid sequence of SEQ ID NO: 2 (e.g., a peptide having at least85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.5%, or 100% sequence identity tothe amino acid sequence of SEQ ID NO: 2). In some embodiments, the aminoacid sequence of the CXCR2 agonist differs from that of SEQ ID NO: 2only by way of one or more conservative amino acid substitutions. Insome embodiments, in some embodiments, the amino acid sequence of theCXCR2 agonist differs from that of SEQ ID NO: 2 by no more than 20, nomore than 15, no more than 10, no more than 5, or no more than 1nonconservative amino acid substitutions.

Additional examples of CXCR2 agonists useful in conjunction with thecompositions and methods described herein are variants of Gro-β N69D,such as peptides that have one or more amino acid substitutions,insertions, and/or deletions relative to Gro-β N69D. In someembodiments, the CXCR2 agonist may be a peptide having at least 85%sequence identity to the amino acid sequence of SEQ ID NO: 3 (e.g., apeptide having at least 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.%, or 100%sequence identity to the amino acid sequence of SEQ ID NO: 3). In someembodiments, the amino acid sequence of the CXCR2 agonist differs fromthat of SEQ ID NO: 3 only by way of one or more conservative amino acidsubstitutions. In some embodiments, in some embodiments, the amino acidsequence of the CXCR2 agonist differs from that of SEQ ID NO: 3 by nomore than 20, no more than 15, no more than 10, no more than 5, or nomore than 1 nonconservative amino acid substitutions.

Additional examples of CXCR2 agonists useful in conjunction with thecompositions and methods described herein are variants of Gro-β T N65D,such as peptides that have one or more amino acid substitutions,insertions, and/or deletions relative to Gro-β T N65D. In someembodiments, the CXCR2 agonist may be a peptide having at least 85%sequence identity to the amino acid sequence of SEQ ID NO: 4 (e.g., apeptide having at least 85%, 90%, 95%, 96%, 97%, 98%, 99%, 99.%, or 100%sequence identity to the amino acid sequence of SEQ ID NO: 4). In someembodiments, the amino acid sequence of the CXCR2 agonist differs fromthat of SEQ ID NO: 4 only by way of one or more conservative amino acidsubstitutions. In some embodiments, in some embodiments, the amino acidsequence of the CXCR2 agonist differs from that of SEQ ID NO: 4 by nomore than 20, no more than 15, no more than 10, no more than 5, or nomore than 1 nonconservative amino acid substitutions.

Agonistic Anti-CXCR2 Antibodies and Antigen-Binding Fragments Thereof

In some embodiments, the CXCR2 agonist is an antibody or antigen-bindingfragment thereof that binds CXCR2 and activates CXCR2 signaltransduction. In some embodiments, the CXCR2 agonist may be an antibodyor antigen-binding fragment thereof that binds the same epitope on CXCR2as Gro-β or a variant or truncation thereof, such as Gro-β T, asassessed, for example, by way of a competitive CXCR2 binding assay. Insome embodiments, the CXCR2 agonist is an antibody or an antigen-bindingfragment thereof that competes with Gro-β or a variant or truncationthereof, such as Gro-β T, for binding to CXCR2.

In some embodiments of any of the above aspects, the antibody orantigen-binding fragment thereof is selected from the group consistingof a monoclonal antibody or antigen-binding fragment thereof, apolyclonal antibody or antigen-binding fragment thereof, a humanizedantibody or antigen-binding fragment thereof, a bispecific antibody orantigen-binding fragment thereof, a dual-variable immunoglobulin domain,a single-chain Fv molecule (scFv), a diabody, a triabody, a nanobody, anantibody-like protein scaffold, a Fv fragment, a Fab fragment, a F(ab′)₂molecule, and a tandem di-scFv. In some embodiments, the antibody has anisotype selected from the group consisting of IgG, IgA, IgM, IgD, andIgE.

Synthetic CXCR2 Agonists

The peptidic CXCR2 agonists described herein, such as Gro-β, Gro-β T,and variants thereof, may be prepared synthetically, for instance, usingsolid phase peptide synthesis techniques. Systems and processes forperforming solid phase peptide synthesis include those that are known inthe art and have been described, for instance, in U.S. Pat. Nos.9,169,287; 9,388,212; 9,206,222; 6,028,172; and 5,233,044, among others,the disclosures of each of which are incorporated herein by reference asthey pertain to protocols and techniques for the synthesis of peptideson solid support. Solid phase peptide synthesis is a process in whichamino acid residues are added to peptides that have been immobilized ona solid support, such as a polymeric resin (e.g., a hydrophilic resin,such as a polyethylene-glycol-containing resin, or hydrophobic resin,such as a polystyrene-based resin).

Peptides, such as those containing protecting groups at amino, hydroxy,thiol, and carboxy substituents, among others, may be bound to a solidsupport such that the peptide is effectively immobilized on the solidsupport. For example, the peptides may be bound to the solid support viatheir C termini, thereby immobilizing the peptides for subsequentreaction in at a resin-liquid interface.

The process of adding amino acid residues to immobilized peptides caninclude exposing a deprotection reagent to the immobilized peptides toremove at least a portion of the protection groups from at least aportion of the immobilized peptides. The deprotection reagent exposurestep can be configured, for instance, such that side-chain protectiongroups are preserved, while N-terminal protection groups are removed.For instance, an exemplary amino protecting contains afluorenylmethyloxycarbonyl (Fmoc) substituent. A deprotection reagentcontaining a strongly basic substance, such as piperidine (e.g., apiperidine solution in an appropriate organic solvent, such as dimethylformamide (DMF)) may be exposed to the immobilized peptides such thatthe Fmoc protecting groups are removed from at least a portion of theimmobilized peptides. Other protecting groups suitable for theprotection of amino substituents include, for instance, thetert-butyloxycarbonyl (Boc) moiety. A deprotection reagent comprising astrong acid, such as trifluoroacetic acid (TFA) may be exposed toimmobilized peptides containing a Boc-protected amino substituent so asto remove the Boc protecting group by an ionization process. In thisway, peptides can be protected and deprotected at specific sites, suchas at one or more side-chains or at the N- or C-terminus of animmobilized peptide so as to append chemical functionalityregioselectively at one or more of these positions. This can be used,for instance, to derivatize a side-chain of an immobilized peptide, orto synthesize a peptide, e.g., from the C-terminus to the N-terminus.

The process of adding amino acid residues to immobilized peptides caninclude, for instance, exposing protected, activated amino acids to theimmobilized peptides such that at least a portion of the activated aminoacids are bonded to the immobilized peptides to form newly-bonded aminoacid residues. For example, the peptides may be exposed to activatedamino acids that react with the deprotected N-termini of the peptides soas to elongate the peptide chain by one amino acid. Amino acids can beactivated for reaction with the deprotected peptides by reaction of theamino acid with an agent that enhances the electrophilicity of thebackbone carbonyl carbon of the amino acid. For example, phosphonium anduronium salts can, in the presence of a tertiary base (e.g.,diisopropylethylamine (DIPEA) and triethylamine (TEA), among others),convert protected amino acids into activated species (for example, BOP,PyBOP, HBTU, and TBTU all generate HOBt esters). Other reagents can beused to help prevent racemization that may be induced in the presence ofa base. These reagents include carbodiimides (for example, DCC or WSCDI)with an added auxiliary nucleophile (for example,1-hydroxy-benzotriazole (HOBO, 1-hydroxy-azabenzotriazole (HOAt), orHOSu) or derivatives thereof. Another reagent that can be utilized toprevent racemization is TBTU. The mixed anhydride method, using isobutylchloroformate, with or without an added auxiliary nucleophile, can alsobe used, as well as the azide method, due to the low racemizationassociated with this reagent. These types of compounds can also increasethe rate of carbodiimide-mediated couplings, as well as preventdehydration of Asn and Gln residues. Typical additional reagents includealso bases such as N,N-diisopropylethylamine (DIPEA), triethylamine(TEA) or N-methylmorpholine (NMM). These reagents are described indetail, for instance, in U.S. Pat. No. 8,546,350, the disclosure ofwhich is incorporated herein in its entirety.

During the recombinant expression and folding of Gro-β and Gro-β T inaqueous solution, a particular C-terminal asparagine residue (Asn69within Gro-β and Asn65 within Gro-β T) is prone to deamidation. Thisprocess effectuates the conversion of the asparagine residue to asparticacid. Without wishing to be bound by any theory, the chemical synthesisof Gro-β and Gro-β T may overcome this problem, for instance, byproviding conditions that reduce the exposure of this asparagine residueto nucleophilic solvent. When prepared synthetically (i.e., chemicallysynthesized), for instance, using, e.g., the solid phase peptidesynthesis techniques described above, synthetic Gro-β, Gro-β T, andvariants thereof that may be used in conjunction with the compositionsand methods described herein may have a purity of, e.g., at least about95% relative to the deamidated versions of these peptides (i.e., containless than 5% of the corresponding deamidated peptide). For instance,synthetic Gro-β, Gro-β T, and variants thereof that may be used inconjunction with the compositions and methods described herein may havea purity of about 95%, 95.5%, 96%, 96.5%, 97%, 97.5%, 98%, 98.5%, 99%,99.5%, 99.6%, 99.7%, 99.8%, 99.9%, 99.99%, or more, relative to thedeamidated versions of these peptides (e.g., the Asn69 deamidatedversion of SEQ ID NO: 1 or the Asn65 deamidated version of SEQ ID NO:2). For instance, synthetic Gro-β, Gro-β T, and variants thereof mayhave, for instance, a purity of from about 95% to about 99.99%, such asa purity of from about 95% to about 99.99%, about 96% to about 99.99%,about 97% to about 99.99%, about 98% to about 99.99%, about 99% to about99.99%, about 99.9% to about 99.99%, about 95% to about 99.5%, about 96%to about 99.5%, about 95% to about 99%, or about 97% to about 99%relative to the deamidated versions of these peptides (e.g., the Asn69deamidated version of SEQ ID NO: 1 or the Asn65 deamidated version ofSEQ ID NO: 2).

CXCR4 Antagonists

Exemplary CXCR4 antagonists for use in conjunction with the compositionsand methods described herein are compounds represented by formula (I)

Z-linker-Z′  (I)

or a pharmaceutically acceptable salt thereof, wherein Z is:

-   -   (i) a cyclic polyamine containing from 9 to 32 ring members,        wherein from 2 to 8 of the ring members are nitrogen atoms        separated from one another by 2 or more carbon atoms; or    -   (ii) an amine represented by formula (IA)

-   -   wherein A includes a monocyclic or bicyclic fused ring system        including at least one nitrogen atom and B is H or a substituent        of from 1 to 20 atoms;

and wherein Z′ is:

-   -   (i) a cyclic polyamine containing from 9 to 32 ring members,        wherein from 2 to 8 of the ring members are nitrogen atoms        separated from one another by 2 or more carbon atoms;    -   (ii) an amine represented by formula (IB)

-   -   wherein A′ includes a monocyclic or bicyclic fused ring system        including at least one nitrogen atom and B′ is H or a        substituent of from 1 to 20 atoms; or    -   (iii) a substituent represented by formula (IC)

—N(R)—(CR₂)_(n)—X  (IC)

-   -   wherein each R is independently H or C₁-C₆ alkyl, n is 1 or 2,        and X is an aryl or heteroaryl group or a mercaptan;        wherein the linker is a bond, optionally substituted alkylene        (e.g., optionally substituted C₁-C₆ alkylene), optionally        substituted heteroalkylene (e.g., optionally substituted C₁-C₆        heteroalkylene), optionally substituted alkenylene (e.g.,        optionally substituted C₂-C₆ alkenylene), optionally substituted        heteroalkenylene (e.g., optionally substituted C₂-C₆        heteroalkenylene), optionally substituted alkynylene (e.g.,        optionally substituted C₂-C₆ alkynylene), optionally substituted        heteroalkynylene (e.g., optionally substituted C₂-C₆        heteroalkynylene), optionally substituted cycloalkylene,        optionally substituted heterocycloalkylene, optionally        substituted arylene, or optionally substituted heteroarylene.

In some embodiments, Z and Z′ may each independently a cyclic polyaminecontaining from 9 to 32 ring members, of which from 2 to 8 are nitrogenatoms separated from one another by 2 or more carbon atoms. In someembodiments, Z and Z′ are identical substituents. As an example, Z maybe a cyclic polyamine including from 10 to 24 ring members. In someembodiments, Z may be a cyclic polyamine that contains 14 ring members.In some embodiments, Z includes 4 nitrogen atoms. In some embodiments, Zis 1,4,8,11-tetraazocyclotetradecane.

In some embodiments, the linker is represented by formula (ID)

wherein ring D is an optionally substituted aryl group, an optionallysubstituted heteroaryl group, an optionally substituted cycloalkylgroup, or an optionally substituted heterocycloalkyl group; andX and Y are each independently optionally substituted alkylene (e.g.,optionally substituted C₁-C₆ alkylene), optionally substitutedheteroalkylene (e.g., optionally substituted C₁-C₆ heteroalkylene),optionally substituted alkenylene (e.g., optionally substituted C₂-C₆alkenylene), optionally substituted heteroalkenylene (e.g., optionallysubstituted C₂-C₆ heteroalkenylene), optionally substituted alkynylene(e.g., optionally substituted C₂-C₆ alkynylene), or optionallysubstituted heteroalkynylene (e.g., optionally substituted C₂-C₆heteroalkynylene).

As an example, the linker may be represented by formula (IE)

wherein ring D is an optionally substituted aryl group, an optionallysubstituted heteroaryl group, an optionally substituted cycloalkylgroup, or an optionally substituted heterocycloalkyl group; andX and Y are each independently optionally substituted alkylene (e.g.,optionally substituted C₁-C₆ alkylene), optionally substitutedheteroalkylene (e.g., optionally substituted C₁-C₆ heteroalkylene),optionally substituted C₂-C₆ alkenylene (e.g., optionally substitutedC₂-C₆ alkenylene), optionally substituted heteroalkenylene (e.g.,optionally substituted C₂-C₆ heteroalkenylene), optionally substitutedalkynylene (e.g., optionally substituted C₂-C₆ alkynylene), oroptionally substituted heteroalkynylene (e.g., optionally substitutedC₂-C₆ heteroalkynylene). In some embodiments, X and Y are eachindependently optionally substituted C₁-C₆ alkylene. In someembodiments, X and Y are identical substituents. In some embodiments, Xand Y may be each be methylene, ethylene, n-propylene, n-butylene,n-pentylene, or n-hexylene groups. In some embodiments, X and Y are eachmethylene groups.

The linker may be, for example, 1,3-phenylene, 2,6-pyridine,3,5-pyridine, 2,5-thiophene, 4,4′-(2,2′-bipyrimidine),2,9-(1,10-phenanthroline), or the like. In some embodiments, the linkeris 1,4-phenylene-bis-(methylene).

CXCR4 antagonists useful in conjunction with the compositions andmethods described herein include plerixafor (also referred to herein as“AMD3100” and “Mozibil”), or a pharmaceutically acceptable salt thereof,represented by formula (II),1,1′-[1,4-phenylenebis(methylene)]-bis-1,4,8,11-tetra-azacyclotetradecane.

Additional CXCR4 antagonists that may be used in conjunction with thecompositions and methods described herein include variants ofplerixafor, such as a compound described in U.S. Pat. No. 5,583,131, thedisclosure of which is incorporated herein by reference as it pertainsto CXCR4 antagonists. In some embodiments, the CXCR4 antagonist may be acompound selected from the group consisting of:1,1′-[1,3-phenylenebis(methylene)]-bis-1,4,8,11-tetra-azacyclotetradecane;1,1′-[1,4-phenylene-bis-(methylene)]-bis-1,4,8,11-tetraazacyclotetradecane;bis-zinc or bis-copper complex of1,1′-[1,4-phenylene-bis-(methylene)]-bis-1,4,8,11-tetraazacyclotetradecane;1,1′-[3,3′-biphenylene-bis-(methylene)]-bis-1,4,8,11-tetraazacyclotetradecane;11,11′-[1,4-phenylene-bis-(methylene)]-bis-1,4,7,11-tetraazacyclotetradecane;1,11′-[1,4-phenylene-bis-(methylene)]-1,4,8,11-tetraazacyclotetradecane-1,4,7,11-tetraazacyclotetradecane;1,1′-[2,6-pyridine-bis-(methylene)]-bis-1,4,8,11-tetraazacyclotetradecane;1,1-[3,5-pyridine-bis-(methylene)]-bis-1,4,8,11-tetraazacyclotetradecane;1,1′-[2,5-thiophene-bis-(methylene)]-bis-1,4,8,11-tetraazacyclotetradecane;1,1′-[4,4′-(2,2′-bipyridine)-bis-(methylene)]-bis-1,4,8,11-tetraazacyclotetradecane;1,1′-[2,9-(1,10-phenanthroline)-bis-(methylene)]-bis-1,4,8,11-tetraazacyclotetradecane;1,1′-[1,3-phenylene-bis-(methylene)]-bis-1,4,7,10-tetraazacyclotetradecane;1,1′-[1,4-phenylene-bis-(methylene)]-bis-1,4,7,10-tetraazacyclotetradecane;1′-[5-nitro-1,3-phenylenebis(methylene)]bis-1,4,8,11-tetraazacyclotetradecane;1′,1′-[2,4,5,6-tetrachloro-1,3-phenylenebis(methylene)]bis-1,4,8,11-tetraazacyclotetradecane;1,1′-[2,3,5,6-tetra-fluoro-1,4-phenylenebis(methylene)]bis-1,4,8,11-tetraazacyclotetradecane;1,1′-[1,4-naphthylene-bis-(methylene)]bis-1,4,8,11-tetraazacyclotetradecane;1,1′-[1,3-phenylenebis-(methylene)]bis-1,5,9-triazacyclododecane;1,1′-[1,4-phenylene-bis-(methylene)]-1,5,9-triazacyclododecane;1,1′-[2,5-dimethyl-1,4-phenylenebis-(methylene)]-bis-1,4,8,11-tetraazacyclotetradecane;1,1′-[2,5-dichloro-1,4-phenylenebis-(methylene)]-bis-1,4,8,11-tetraazacyclotetradecane;1,1′-[2-bromo-1,4-phenylenebis-(methylene)]-bis-1,4,8,11-tetraazacyclotetradecane;and1,1′-[6-phenyl-2,4-pyridinebis-(methylene)]-bis-1,4,8,11-tetraazacyclotetradecane.

In some embodiments, the CXCR4 antagonist is a compound described in US2006/0035829, the disclosure of which is incorporated herein byreference as it pertains to CXCR4 antagonists. In some embodiments, theCXCR4 antagonist may be a compound selected from the group consistingof: 3,7,11,17-tetraazabicyclo(13.3.1)heptadeca-1(17),13,15-triene;4,7,10,17-tetrambicyclo(13.3.1)heptadeca-1(17),13,15-triene;1,4,7,10-tetraazacyclotetradecane; 1,4,7-triazacyclotetradecane; and4,7,10-triazabicyclo(13.3.1)heptadeca-1(17),13,15-triene.

The CXCR4 antagonist may be a compound described in WO 2001/044229, thedisclosure of which is incorporated herein by reference as it pertainsto CXCR4 antagonists. In some embodiments, the CXCR4 antagonist may be acompound selected from the group consisting of:N-[4-(11-fluoro-1,4,7-triazacyclotetradecanyl)-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine;N-[4-(11,11-difluoro-1,4,7-triazacyclotetradecanyl)-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine;N-[4-(1,4,7-triazacyclotetradecan-2-onyl)-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine;N-[12-(5-oxa-1,9-diazacyclotetradecanyl)-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine;N-[4-(11-oxa-1,4,7-triazacyclotetradecanyl)-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine;N-[4-(11-thia-1,4,7-triazacyclotetradecanyl)-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine;N-[4-(11-sulfoxo-1,4,7-triazacyclotetradecanyl)-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine;N-[4-(11-sulfono-1,4,7-triazacyclotetradecanyl)-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine;andN-[4-(3-carboxo-1,4,7-triazacyclotetradecanyl)-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine.

Additional CXCR4 antagonists useful in conjunction with the compositionsand methods described herein include compounds described in WO2000/002870, the disclosure of which is incorporated herein by referenceas it pertains to CXCR4 antagonists. In some embodiments, the CXCR4antagonist may be a compound selected from the group consisting of:N-[1,4,8,11-tetraazacyclotetra-decanyl-1,4-phenylenebis-(methylene)]-2-(aminomethyl)pyridine;N-[1,4,8,11-tetraazacyclotetra-decanyl-1,4-phenylenebis(methylene)]-N-methyl-2-(aminomethyl)pyridine;N-[1,4,8,11-tetraazacyclotetra-decanyl-1,4-phenylenebis(methylene)]-4-(aminomethyl)pyridine;N-[1,4,8,11-tetraazacyclotetra-decanyl-1,4-phenylenebis(methylene)]-3-(aminomethyl)pyridine;N-[1,4,8,11-tetraazacyclotetra-decanyl-1,4-phenylenebis(methylene)]-(2-aminomethyl-5-methyl)pyrazine;N-[1,4,8,11-tetraazacyclotetra-decanyl-1,4-phenylenebis(methylene)]-2-(aminoethyl)pyridine;N-[1,4,8,11-tetraazacyclotetra-decanyl-1,4-phenylenebis(methylene)]-2-(aminomethyl)thiophene;N-[1,4,8,11-tetraazacyclotetra-decanyl-1,4-phenylenebis(methylene)]-2-(aminomethyl)mercaptan;N-[1,4,8,11-tetraazacyclotetra-decanyl-1,4-phenylenebis(methylene)]-2-aminobenzylamine;N-[1,4,8,11-tetraazacyclotetra-decanyl-1,4-phenylenebis(methylene)]-4-aminobenzylamine;N-[1,4,8,11-tetraazacyclotetra-decanyl-1,4-phenylenebis(methylene)]-4-(aminoethyl)imidazole;N-[1,4,8,11-tetraazacyclotetra-decanyl-1,4-phenylenebis(methylene)]-benzylamine;N-[4-(1,4,7-triazacyclotetra-decanyl)-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine;N-[7-(4,7,10,17-tetraazabicyclo[13.3.1]heptadeca-1(17),13,15-trienyl)-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine;N-[7-(4,7,10-triazabicyclo[13.3.1]heptadeca-1(17),13,15-trienyl)-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine;N-[1-(1,4,7-triazacyclotetra-decanyl)-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine;N-[4-[4,7,10,17-tetraazabicyclo[13.3.1]heptadeca-1(17),13,15-trienyl]-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine;N-[4-[4,7,10-triazabicyclo[13.3.1]heptadeca-1(17),13,15-trienyl]-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine;N-[1,4,8,11-tetraazacyclotetradecanyl-1,4-phenylenebis(methylene)]-purine;1-[1,4,8,11-tetraazacyclotetradecanyl-1,4-phenylenebis(methylene)]-4-phenylpiperazine;N-[4-(1,7-diazacyclotetradecanyl)-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine;andN-[7-(4,10-diazabicyclo[13.3.1]heptadeca-1(17),13,15-trienyl)-1,4-phenylenebis(methylene)]-2-(aminomethyl)pyridine.

In some embodiments, the CXCR4 antagonist is a compound selected fromthe group consisting of:1-[2,6-dimethoxypyrid-4-yl(methylene)]-1,4,8,11-tetraazacyclotetradecane;1-[2-chloropyrid-4-yl(methylene)]-1,4,8,11-tetraazacyclotetradecane;1-[2,6-dimethylpyrid-4-yl(methylene)]-1,4,8,11-tetraazacyclotetradecane;1-[2-methylpyrid-4-yl(methylene)]-1,4,8,11-tetraazacyclotetradecane;1-[2,6-dichloropyrid-4-yl(methylene)]-1,4,8,11-tetraazacyclotetradecane;1-[2-chloropyrid-5-yl(methylene)]-1,4,8,11-tetraazacyclotetradecane; and7-[4-methylphenyl(methylene)]-4,7,10,17-tetraazabicyclo[13.3.1]heptadeca-1(17),13,15-triene.

In some embodiments, the CXCR4 antagonist is a compound described inU.S. Pat. No. 5,698,546, the disclosure of which is incorporated hereinby reference as it pertains to CXCR4 antagonists. In some embodiments,the CXCR4 antagonist may be a compound selected from the groupconsisting of:7,7′-[1,4-phenylene-bis(methylene)]bis-3,7,11,17-tetraazabicyclo[13.3.1]heptadeca-1(17),13,15-triene;7,7′-[1,4-phenylene-bis(methylene)]bis[15-chloro-3,7,11,17-tetraazabicyclo[13.3.1]heptadeca-1(17),13,15-triene];7,7′-[1,4-phenylene-bis(methylene)]bis[15-methoxy-3,7,11,17-tetraazabicyclo[13.3.1]heptadeca-1(17),13,15-triene];7,7′-[1,4-phenylene-bis(methylene)]bis-3,7,11,17-tetraazabicyclo[13.3.1]-heptadeca-13,16-triene-15-one;7,7′-[1,4-phenylene-bis(methylene)]bis-4,7,10,17-tetraazabicyclo[13.3.1]-heptadeca-1(17),13,15-triene;8,8′-[1,4-phenylene-bis(methylene)]bis-4,8,12,19-tetraazabicyclo[15.3.1]nonadeca-1(19),15,17-triene;6,6′-[1,4-phenylene-bis(methylene)]bis-3,6,9,15-tetraazabicyclo[11.3.1]pentadeca-1(15),11,13-triene;6,6′-[1,3-phenylene-bis(methylene)]bis-3,6,9,15-tetraazabicyclo[11.3.1]pentadeca-1(15),11,13-triene; and17,17′-[1,4-phenylene-bis(methylene)]bis-3,6,14,17,23,24-hexaazatricyclo[17.3.1.1⁸′¹²]tetracosa-1(23),8,10,12(24),19,21-hexaene.

In some embodiments, the CXCR4 antagonist is a compound described inU.S. Pat. No. 5,021,409, the disclosure of which is incorporated hereinby reference as it pertains to CXCR4 antagonists. In some embodiments,the CXCR4 antagonist may be a compound selected from the groupconsisting of: 2,2′-bicyclam, 6,6′-bicyclam; 3,3′-(bis-1,5,9,13-tetraazacyclohexadecane); 3,3′-(bis-1,5,8,11,14-pentaazacyclohexadecane);methylene (or polymethylene) di-1-N-1,4,8,11-tetraazacyclotetradecane;3,3′-bis-1,5,9,13-tetraazacyclohexadecane;3,3′-bis-1,5,8,11,14-pentaazacyclohexadecane;5,5′-bis-1,4,8,11-tetraazacyclotetradecane;2,5′-bis-1,4,8,11-tetraazacyclotetradecane;2,6′-bis-1,4,8,11-tetraazacyclotetradecane;11,11′-(1,2-ethanediyl)bis-1,4,8,11-tetraazacyclotetradecane;11,11′-(1,2-propanediyl)bis-1,4,8,11-tetraazacyclotetradecane;11,11′-(1,2-butanediyl)bis-1,4,8,11-tetraazacyclotetradecane;11,11′-(1,2-pentanediyl)bis-1,4,8,11-tetraazacyclotetradecane; and11,11′-(1,2-hexanediyl)bis-1,4,8,11-tetraazacyclotetradecane.

In some embodiments, the CXCR4 antagonist is a compound described in WO2000/056729, the disclosure of which is incorporated herein by referenceas it pertains to CXCR4 antagonists. In some embodiments, the CXCR4antagonist may be a compound selected from the group consisting of:N-(2-pyridinylmethyl)-N′-(6,7,8,9-tetrahydro-5H-cyclohepta[b]pyridin-9-yl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-(6,7-dihydro-5H-cyclopenta[b]pyridin-7-yl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-(1,2,3,4-tetrahydro-1-naphthalenyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-(1-naphthalenyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-(8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-[2-[(2-pyridinylmethyl)amino]ethyl]-N′-(1-methyl-1,2,3,4-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-[2-[(1H-imidazol-2-ylmethyl)amino]ethyl]-N′-(1-methyl-1,2,3,4-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-(1,2,3,4-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-[2-[(1H-imidazol-2-ylmethyl)amino]ethyl]-N′-(1,2,3,4-tetrahydro-1-naphthalenyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-(2-phenyl-5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N,N′-bis(2-pyridinylmethyl)-N′-(2-phenyl-5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;

N-(2-pyridinylmethyl)-N′-(5,6,7,8-tetrahydro-5-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-(1H-imidazol-2-ylmethyl)-N′-(5,6,7,8-tetrahydro-5-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-(1H-imidazol-2-ylmethyl)-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-[(2-amino-3-phenyl)propyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-(1H-imidazol-4-ylmethyl)-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-(2-quinolinylmethyl)-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-(2-(2-naphthoyl)aminoethyl)-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′—[(S)-(2-acetylamino-3-phenyl)propyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′—[(S)-(2-acetylamino-3-phenyl)propyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-[3-((2-naphthalenylmethyl)amino)propyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-[2-(S)-pyrollidinylmethyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-[2-(R)-pyrollidinylmethyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-[3-pyrazolylmethyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-[2-pyrrolylmethyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-[2-thiopheneylmethyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-[2-thiazolylmethyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-[2-furanylmethyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-[2-[(phenylmethyl)amino]ethyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-(2-aminoethyl)-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-3-pyrrolidinyl-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-4-piperidinyl-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-[2-[(phenyl)amino]ethyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-(7-methoxy-1,2,3,4-tetrahydro-2-naphthalenyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-(6-methoxy-1,2,3,4-tetrahydro-2-naphthalenyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-(1-methyl-1,2,3,4-tetrahydro-2-naphthalenyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-(7-methoxy-3,4-dihydronaphthalenyl)-1-(aminomethyl)-4-benzamide;N-(2-pyridinylmethyl)-N′-(6-methoxy-3,4-dihydronaphthalenyl)-1-(aminomethyl)-4-benzamide;N-(2-pyridinylmethyl)-N′-(1H-imidazol-2-ylmethyl)-N′-(7-methoxy-1,2,3,4-tetrahydro-2-naphthalenyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-(8-hydroxy-1,2,3,4-tetrahydro-2-naphthalenyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-(1H-imidazol-2-ylmethyl)-N′-(8-hydroxy-1,2,3,4-tetrahydro-2-naphthalenyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-(8-Fluoro-1,2,3,4-tetrahydro-2-naphthalenyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-(1H-imidazol-2-ylmethyl)-N′-(8-Fluoro-1,2,3,4-tetrahydro-2-naphthalenyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-(5,6,7,8-tetrahydro-7-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-(1H-imidazol-2-ylmethyl)-N′-(5,6,7,8-tetrahydro-7-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-[2-[(2-naphthalenylmethyl)amino]ethyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-[2-(isobutylamino)ethyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-[2-[(2-pyridinylmethyl)amino]ethyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-[2-[(2-furanylmethyl)amino]ethyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-(2-guanidinoethyl)-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-[2-[bis-[(2-methoxy)phenylmethyl]amino]ethyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-[2-[(1H-imidazol-4-ylmethyl)amino]ethyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-[2-[(1H-imidazol-2-ylmethyl)amino]ethyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-[2-(phenylureido)ethyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′—[[N″-(n-butyl)carboxamido]methyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-(carboxamidomethyl)-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′—[(N″-phenyl)carboxamidomethyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-(carboxymethyl)-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-(phenylmethyl)-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-(1H-benzimidazol-2-ylmethyl)-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-(5,6-dimethyl-1H-benzimidazol-2-ylmethyl)-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine(hydrobromide salt);N-(2-pyridinylmethyl)-N′-(5-nitro-1H-benzimidazol-2-ylmethyl)-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-[(1H)-5-azabenzimidazol-2-ylmethyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N-(4-phenyl-1H-imidazol-2-ylmethyl)-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-[2-(2-pyridinyl)ethyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-(2-benzoxazolyl)-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-(trans-2-aminocyclohexyl)-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-(2-phenylethyl)-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-(3-phenylpropyl)-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N′-(trans-2-aminocyclopentyl)-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-[[4-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]-N-(5,6,7,8-tetrahydro-8-quinolinyl)-glycinamide;N-[[4-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]-N-(5,6,7,8-tetrahydro-8-quinolinyl)-(L)-alaninamide;N-[[4-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]-N-(5,6,7,8-tetrahydro-8-quinolinyl)-(L)-aspartamide;N-[[4-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]-N-(5,6,7,8-tetrahydro-8-quinolinyl)-pyrazinamide;N-[[4-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]-N-(5,6,7,8-tetrahydro-8-quinolinyl)-(L)-prolinamide;N-[[4-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]-N-(5,6,7,8-tetrahydro-8-quinolinyl)-(L)-lysinamide;N-[[4-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]-N-(5,6,7,8-tetrahydro-8-quinolinyl)-benzamide;N-[[4-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]-N-(5,6,7,8-tetrahydro-8-quinolinyl)-picolinamide;N′-Benzyl-N-[[4-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]-N-(5,6,7,8-tetrahydro-8-quinolinyl)-urea;N′-phenyl-N-[[4-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]-N-(5,6,7,8-tetrahydro-8-quinolinyl)-urea;N-(6,7,8,9-tetrahydro-5H-cyclohepta[bacteriapyridin-9-yl)-44[(2-pyridinylmethyl)amino]methyl]benzamide;N-(5,6,7,8-tetrahydro-8-quinolinyl)-4-[[(2-pyridinylmethyl)amino]methyl]benzamide;N,N′-bis(2-pyridinylmethyl)-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N,N′-bis(2-pyridinylmethyl)-N′-(6,7,8,9-tetrahydro-5H-cyclohepta[bacteriapyridin-9-yl)-1,4-benzenedimethanamine;N,N′-bis(2-pyridinylmethyl)-N′-(6,7-dihydro-5H-cyclopenta[bacteriapyridin-7-yl)-1,4-benzenedimethanamine;N,N′-bis(2-pyridinylmethyl)-N′-(1,2,3,4-tetrahydro-1-naphthalenyl)-1,4-benzenedimethanamine;N,N′-bis(2-pyridinylmethyl)-N′-[(5,6,7,8-tetrahydro-8-quinolinyl)methyl]-1,4-benzenedimethanamine;N,N′-bis(2-pyridinylmethyl)-N′[(6,7-dihydro-5H-cyclopenta[bacteriapyridin-7-yl)methyl]-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N-(2-methoxyethyl)-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(2-pyridinylmethyl)-N-[2-(4-methoxyphenyl)ethyl]-N′-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N,N′-bis(2-pyridinylmethyl)-1,4-(5,6,7,8-tetrahydro-8-quinolinyl)benzenedimethanamine;N-[(2,3-dimethoxyphenyl)methyl]-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N,N′-bis(2-pyridinylmethyl)-N-[1-(N″-phenyl-N″-methylureido)-4-piperidinyl]-1,3-benzenedimethanamine;N,N′-bis(2-pyridinylmethyl)-N—[N″-p-toluenesulfonylphenylalanyl)-4-piperidinyl]-1,3-benzenedimethanamine;N,N′-bis(2-pyridinylmethyl)-N-[1-[3-(2-chlorophenyl)-5-methyl-isoxazol-4-oyl]-4-piperidinyl]-1,3-benzenedimethanamine;N-[(2-hydroxyphenyl)methyl]-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-cyclohepta[bacteriapyridin-9-yl)-1,4-benzenedimethanamine;N-[(4-cyanophenyl)methyl]-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-cyclohepta[bacteriapyridin-9-yl)-1,4-benzenedimethanamine;N-[(4-cyanophenyl)methyl]-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-[(4-acetamidophenyl)methyl]-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-[(4-phenoxyphenyl)methyl]-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-cyclohepta[bacteriapyridin-9-yl)-1,4-benzenedimethanamine;N-[(1-methyl-2-carboxamido)ethyl]-N,N′-bis(2-pyridinylmethyl)-1,3-benzenedimethanamine;N-[(4-benzyloxyphenyl)methyl]-M-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-cyclohepta[bacteriapyridin-9-yl)-1,4-benzenedimethanamine;N-[(thiophene-2-yl)methyl]-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-cyclohepta[bacteriapyridin-9-yl)-1,4-benzenedimethanamine;N-[1-(benzyl)-3-pyrrolidinyl]-N,N′-bis(2-pyridinylmethyl)-1,3-benzenedimethanamine;N-[[1-methyl-3-(pyrazol-3-yl)]propyl]-N,N′-bis(2-pyridinylmethyl)-1,3-benzenedimethanamine;N-[1-(phenyl)ethyl]-N,N′-bis(2-pyridinylmethyl)-1,3-benzenedimethanamine;N-[(3,4-methylenedioxyphenyl)methyl]-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-cyclohepta[b]pyridin-9-yl)-1,4-benzenedimethanamine;N-[1-benzyl-3-carboxymethyl-4-piperidinyl]-N,N′-bis(2-pyridinylmethyl)-1,3-benzenedimethanamine;N-[(3,4-methylenedioxyphenyl)methyl]-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(3-pyridinylmethyl)-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-cyclohepta[b]pyridin-9-yl)-1,4-benzenedimethanamine;N-[[1-methyl-2-(2-tolyl)carboxamido]ethyl]-N,N′-bis(2-pyridinylmethyl)-1,3-benzenedimethanamine;N-[(1,5-dimethyl-2-phenyl-3-pyrazolinone-4-yl)methyl]-M-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-[(4-propoxyphenyl)methyl]-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-cyclohepta[b]pyridin-9-yl)-1,4-benzenedimethanamine;N-(1-phenyl-3,5-dimethylpyrazolin-4-ylmethyl)-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N—[H-imidazol-4-ylmethyl]-N,N′-bis(2-pyridinylmethyl)-1,3-benzenedimethanamine;N-[(3-methoxy-4,5-methylenedioxyphenyl)methyl]-N′-[2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-cyclohepta[b]pyridin-9-yl)-1,4-benzenedimethanamine;N-[(3-cyanophenyl)methyl]-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-cyclohepta[b]pyridin-9-yl)-1,4-benzenedimethanamine;N-[(3-cyanophenyl)methyl]-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(5-ethylthiophene-2-ylmethyl)-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-cyclohepta[b]pyridin-9-yl)-1,4-benzenedimethanamine;N-(5-ethylthiophene-2-ylmethyl)-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-[(2,6-difluorophenyl)methyl]-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-cyclohepta[b]pyridin-9-yl)-1,4-benzenedimethanamine;N-[(2,6-difluorophenyl)methyl]-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-[(2-difluoromethoxyphenyl)methyl]-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-cyclohepta[b]pyridin-9-yl)-1,4-benzenedimethanamine;N-(2-difluoromethoxyphenylmethyl)-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(1,4-benzodioxan-6-ylmethyl)-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-cyclohepta[b]pyridin-9-yl)-1,4-benzenedimethanamine;N,N′-bis(2-pyridinylmethyl)-N-[1-(N″-phenyl-N″-methylureido)-4-piperidinyl]-1,4-benzenedimethanamine;N,N′-bis(2-pyridinylmethyl)-N—[N″-p-toluenesulfonylphenylalanyl)-4-piperidinyl]-1,4-benzenedimethanamine;N-[1-(3-pyridinecarboxamido)-4-piperidinyl]-N,N′-bis(2-pyridinylmethyl)-1,4-benzenedimethanamine;N-[1-(cyclopropylcarboxamido)-4-piperidinyl]-N,N′-bis(2-pyridinylmethyl)-1,4-benzenedimethanamine;N-[1-(1-phenylcyclopropylcarboxamido)-4-piperidinyl]-N,N′-bis(2-pyridinylmethyl)-1,4-benzenedimethanamine;N-(1,4-benzodioxan-6-ylmethyl)-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-[1-[3-(2-chlorophenyl)-5-methyl-isoxazol-4-carboxamido]-4-piperidinyl]-N,N′-bis(2-pyridinylmethyl)-1,4-benzenedimethanamine;N-[1-(2-thiomethylpyridine-3-carboxamido)-4-piperidinyl]-N,N′-bis(2-pyridinylmethyl)-1,4-benzenedimethanamine;N-[(2,4-difluorophenyl)methyl]-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(1-methylpyrrol-2-ylmethyl)-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-[(2-hydroxyphenyl)methyl]-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-[(3-methoxy-4,5-methylenedioxyphenyl)methyl]-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(3-pyridinylmethyl)-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-[2-(N″-morpholinomethyl)-1-cyclopentyl]-N,N′-bis(2-pyridinylmethyl)-1,4-benzenedimethanamine;N-[(1-methyl-3-piperidinyl)propyl]-N,N′-bis(2-pyridinylmethyl)-1,4-benzenedimethanamine;N-(1-methylbenzimidazol-2-ylmethyl)-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-[1-(benzyl)-3-pyrrolidinyl]-N,N′-bis(2-pyridinylmethyl)-1,4-benzenedimethanamine;N-[[(1-phenyl-3-(N″-morpholino)]propyl]-N,N′-bis(2-pyridinylmethyl)-1,4-benzenedimethanamine;N-[1-(iso-propyl)-4-piperidinyl]-N,N′-bis(2-pyridinylmethyl)-1,4-benzenedimethanamine;N-[1-(ethoxycarbonyl)-4-piperidinyl]-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-[(1-methyl-3-pyrazolyl)propyl]-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-[1-methyl-2-(N″,N″-diethylcarboxamido)ethyl]-N,N′-bis(2-pyridinylmethyl)-1,4-benzenedimethanamine;N-[(1-methyl-2-phenylsulfonyl)ethyl]-M-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-[(2-chloro-4,5-methylenedioxyphenyl)methyl]-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-[1-methyl-2-[N″-(4-chlorophenyl)carboxamido]ethyl]-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(1-acetoxyindol-3-ylmethyl)-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-cyclohepta[b]pyridin-9-yl)-1,4-benzenedimethanamine;N-[(3-benzyloxy-4-methoxyphenyl)methyl]-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-cyclohepta[b]pyridin-9-yl)-1,4-benzenedimethanamine;N-(3-quinolylmethyl)-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-[(8-hydroxy)-2-quinolylmethyl]-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-cyclohepta[b]pyridin-9-yl)-1,4-benzenedimethanamine;N-(2-quinolylmethyl)-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-cyclohepta[b]pyridin-9-yl)-1,4-benzenedimethanamine;N-[(4-acetamidophenyl)methyl]-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-cyclohepta[b]pyridin-9-yl)-1,4-benzenedimethanamine;N-[1H-imidazol-2-ylmethyl]-N,N′-bis(2-pyridinylmethyl)-1,4-benzenedimethanamine;N-(3-quinolylmethyl)-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-cyclohepta[b]pyridin-9-yl)-1,4-benzenedimethanamine;N-(2-thiazolylmethyl)-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-cyclohepta[b]pyridin-9-yl)-1,4-benzenedimethanamine;N-(4-pyridinylmethyl)-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-cyclohepta[b]pyridin-9-yl)-1,4-benzenedimethanamine;N-[(5-benzyloxy)benzo[b]pyrrol-3-ylmethyl]-N,N′-bis(2-pyridinylmethyl)-1,4-benzenedimethanamine;N-(1-methylpyrazol-2-ylmethyl)-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-cyclohepta[b]pyridin-9-yl)-1,4-benzenedimethanamine;N-[(4-methyl)-1H-imidazol-5-ylmethyl]-N,N′-bis(2-pyridinylmethyl)-1,4-benzenedimethanamine;N-[[(4-dimethylamino)-1-napthalenyl]methyl]-N,N′-bis(2-pyridinylmethyl)-1,4-benzenedimethanamine;N-[1,5-dimethyl-2-phenyl-3-pyrazolinone-4-ylmethyl]-N,N′-bis(2-pyridinylmethyl)-1,4-benzenedimethanamine;N-[1-[(1-acetyl-2-(R)-prolinyl]-4-piperidinyl]-N-[2-(2-pyridinyl)ethyl]-N′-(2-pyridinylmethyl)-1,3-benzenedimethanamine;N-[1-[2-acetamidobenzoyl-4-piperidinyl]-4-piperidinyl]-N-[2-(2-pyridinyl)ethyl]-N′-(2-pyridinylmethyl)-1,3-benzenedimethanamine;N-[(2-cyano-2-phenyl)ethyl]-M-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-cyclohepta[b]pyridin-9-yl)-1,4-benzenedimethanamine;N—[(N″-acetyltryptophanyl)-4-piperidinyl]-N-[2-(2-pyridinyl)ethyl]-N′-(2-pyridinylmethyl)-1,3-benzenedimethanamine;N—[(N″-benzoylvalinyl)-4-piperidinyl]-N-[2-(2-pyridinyl)ethyl]-N′-(2-pyridinylmethyl)-1,3-benzenedimethanamine;N-[(4-dimethylaminophenyl)methyl]-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-cyclohepta[b]pyridin-9-yl)-1,4-benzenedimethanamine;N-(4-pyridinylmethyl)-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(1-methylbenzimadazol-2-ylmethyl)-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-cyclohepta[b]pyridin-9-yl)-1,4-benzenedimethanamine;N-[1-butyl-4-piperidinyl]-N-[2-(2-pyridinyl)ethyl]-N′-(2-pyridinylmethyl)-1,3-benzenedimethanamine;N-[1-benzoyl-4-piperidinyl]-N-[2-(2-pyridinyl)ethyl]-N′-(2-pyridinylmethyl)-1,3-benzenedimethanamine;N-[1-(benzyl)-3-pyrrolidinyl]-N-[2-(2-pyridinyl)ethyl]-N′-(2-pyridinylmethyl)-1,3-benzenedimethanamine;N-[(1-methyl)benzo[b]pyrrol-3-ylmethyl]-N-[2-(2-pyridinyl)ethyl]-N′-(2-pyridinylmethyl)-1,3-benzenedimethanamine;N-[1H-imidazol-4-ylmethyl]-N-[2-(2-pyridinyl)ethyl]-N′-(2-pyridinylmethyl)-1,3-benzenedimethanamine;N-[1-(benzyl)-4-piperidinyl]-N-[2-(2-pyridinyl)ethyl]-N′-(2-pyridinylmethyl)-1,4-benzenedimethanamine;N-[1-methylbenzimidazol-2-ylmethyl]-N-[2-(2-pyridinyl)ethyl]-N′-(2-pyridinylmethyl)-1,4-benzenedimethanamine;N-[(2-phenyl)benzo[b]pyrrol-3-ylmethyl]-N-[2-(2-pyridinyl)ethyl]-N′-(2-pyridinylmethyl)-1,4-benzenedimethanamine;N-[(6-methylpyridin-2-yl)methyl]-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,4-benzenedimethanamine;N-(3-methyl-1H-pyrazol-5-ylmethyl)-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,3-benzenedimethanamine;N-[(2-methoxyphenyl)methyl]-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,3-benzenedimethanamine;N-[(2-ethoxyphenyl)methyl]-N′-(2-pyridinylmethyl)-N-(6,7,8,9-tetrahydro-5H-cyclohepta[b]pyridin-9-yl)-1,3-benzenedimethanamine;N-(benzyloxyethyl)-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,3-benzenedimethanamine;N-[(2-ethoxy-1-naphthalenyl)methyl]-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,3-benzenedimethanamine;N-[(6-methylpyridin-2-yl)methyl]-N′-(2-pyridinylmethyl)-N-(5,6,7,8-tetrahydro-8-quinolinyl)-1,3-benzenedimethanamine;1-[[4-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]guanidine;N-(2-pyridinylmethyl)-N-(8-methyl-8-azabicyclo[3.2.1]octan-3-yl)-1,4-benzenedimethanamine;1-[[4-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]homopiperazine;1-[[3-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]homopiperazine;trans andcis-1-[[4-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]-3,5-piperidinediamine;N,N′-[1,4-Phenylenebis(methylene)]bis-4-(2-pyrimidyl)piperazine;1-[[4-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]-1-(2-pyridinyl)methylamine;2-(2-pyridinyl)-5-[[(2-pyridinylmethyl)amino]methyl]-1,2,3,4-tetrahydroisoquinoline;1-[[4-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]-3,4-diaminopyrrolidine;1-[[4-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]-3,4-diacetylaminopyrrolidine;[[4-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]-2,5,8-triaza-3-oxabicyclo[4.3.0]nonane;and8-[[4-[[(2-pyridinylmethyl)amino]methyl]phenyl]methyl]-2,5,8-triazabicyclo[4.3.0]nonane.

Additional CXCR4 antagonists that may be used to in conjunction with thecompositions and methods described herein include those described in WO2001/085196, WO 1999/050461, WO 2001/094420, and WO 2003/090512, thedisclosures of each of which are incorporated herein by reference asthey pertain to compounds that inhibit CXCR4 activity or expression.

Expansion of Hematopoietic Stem and Progenitor Cells

Prior to infusion into a patient, hematopoietic and progenitor cells maybe expanded ex vivo, for example, by contacting the cells with an arylhydrocarbon receptor antagonist. Aryl hydrocarbon receptor antagonistsuseful in conjunction with the compositions and methods described hereininclude those described in U.S. Pat. No. 9,580,426, the disclosure ofwhich is incorporated herein by reference in its entirety.

In some embodiments, aryl hydrocarbon receptor antagonists include thoserepresented by formula (III)

in which:L is selected from —NR_(5a)(CH₂)₂₋₃, —NR_(5a)(CH₂)₂NR_(5b)—,—NR_(5a)(CH₂)₂S—, —NR_(5a)CH₂CH(OH)— and —NR_(5a)CH(CH₃)CH₂—; whereinR_(5a) and R_(5b) are independently selected from hydrogen and C₁₋₄alkyl;R₁ is selected from thiophenyl, 1H-benzoimidazolyl, isoquinolinyl,1H-imidazopyridinyl, benzothiophenyl, pyrimidinyl, pyridinyl, pyrazinyl,pyridazinyl, and thiazolyl; In some embodiments, wherein the thiophenyl,1H-benzoimidazolyl, isoquinolinyl, 1H-imidazopyridinyl, benzothiophenyl,pyrimidinyl, pyridinyl, pyrazinyl, pyridazinyl, or thiazolyl of R₁ canbe optionally substituted by 1 to 3 radicals independently selected fromcyano, hydroxy, C₁₋₄ alkyl, C₁₋₄ alkoxy, halo, halo-substituted-C₁₋₄alkyl, halo-substituted-C₁₋₄alkoxy, amino, —C(O)R_(8a), —S(O)₀₋₂R_(8a),—C(O)OR_(8a) and —C(O)NR_(8a)R_(8b); wherein R_(8a) and R_(8b) areindependently selected from hydrogen and C₁₋₄alkyl;R₂ is selected from —S(O)₂NR_(6a)R_(6b), —NR_(6a)C(O)R_(6b)—,—NR_(6a)C(O)NR_(6b)R_(6c), phenyl, 1H-pyrrolopyridin-3-yl,1H-pyrrolopyridin-5-yl, 1H-indolyl thiophenyl, pyridinyl,1H-1,2,4-triazolyl, 2-oxoimidazolidinyl, 1H-pyrazolyl,2-oxo-2,3-dihydro-1H-benzoimidazolyl and 1H-indazolyl; wherein R_(6a),R_(6b) and R_(6c) are independently selected from hydrogen andC₁₋₄alkyl; and the phenyl, 1H-pyrrolopyridin-3-yl,1H-pyrrolo[2,3-b]pyridin-5-yl, 1H-indolyl, thiophenyl, pyridinyl,1H-1,2,4-triazolyl, 2-oxoimidazolidinyl, 1H-pyrazolyl,2-oxo-2,3-dihydro-1H-benzoimidazolyl or 1H-indazolyl of R₂ is optionallysubstituted with 1 to 3 radicals independently selected from hydroxy,halo, methyl, methoxy, amino, —O(CH₂)₂NR_(7a)R_(7b),—S(O)₂NR_(7a)R_(7b), —OS(O)₂NR_(7a)R_(b) and —NR_(7a)S(O)₂R_(7b);wherein R_(7a) andR_(7b) are independently selected from hydrogen and C₁₋₄ alkyl;R₃ is selected from hydrogen, C₁₋₄alkyl and biphenyl; andR₄ is selected from C₁₋₁₀ alkyl, prop-1-en-2-yl, cyclohexyl,cyclopropyl, 2-(2-oxopyrrolidin-1-yl)ethyl, oxetan-2-yl, oxetan-3-yl,benzhydryl, tetrahydro-2H-pyran-2-yl, tetrahydro-2H-pyran-3-yl, phenyl,tetrahydrofuran-3-yl, and benzyl, (4-pentylphenyl)(phenyl)methyl and1-(1-(2-oxo-6,9,12-trioxa-3-azatetradecan-14-yl)-1H-1,2,3-triazol-4-yl)ethylwherein said alkyl, cyclopropyl, cyclohexyl,2-(2-oxopyrrolidin-1-yl)ethyl, oxetan-3-yl, oxetan-2-yl, benzhydryl,tetrahydro-2H-pyran-2-yl, tetrahydro-2H-pyran-3-yl, phenyl,tetrahydrofuran-3-yl, benzyl, (4-pentylphenyl)(phenyl)methyl or1-(1-(2-oxo-6,9,12-trioxa-3-azatetradecan-14-yl)-1H-1,2,3-triazol-4-yl)ethylcan be optionally substituted with 1 to 3 radicals independentlyselected from hydroxy, C₁₋₄alkyl and halo-substituted-C₁₋₄alkyl; or asalt thereof.

In some embodiments, aryl hydrocarbon receptor antagonists useful inconjunction with the compositions and methods described herein includeSR-1, represented by formula (1), below.

Methods for the Recombinant Expression of Peptides and Proteins

Peptides and proteins described herein (e.g., CXCR2 agonists, such asGro-β, Gro-β T, Gro-β N69D, T N65D, and variants thereof) can beexpressed in host cells, for example, by delivering to the host cell anucleic acid encoding the corresponding peptide or protein. The sectionsthat follow describe a variety of techniques that can be used for thepurposes of introducing nucleic acids encoding peptides and proteinsdescribed herein to a host cell for the purposes of recombinantexpression.

Transfection Techniques

Techniques that can be used to introduce a polynucleotide, such asnucleic acid encoding a CXCR2 agonist, such as Gro-β, Gro-β T, Gro-βN69D, Gro-β T N65D, or a variant thereof, into a cell (e.g., a mammaliancell, such as a human cell) are known in the art. In some embodiments,electroporation can be used to permeabilize mammalian cells (e.g., humancells) by the application of an electrostatic potential to the cell ofinterest. Mammalian cells, such as human cells, subjected to an externalelectric field in this manner are subsequently predisposed to the uptakeof exogenous nucleic acids. Electroporation of mammalian cells isdescribed in detail, e.g., in Chu et al. (1987) Nucleic Acids Research15:1311, the disclosure of which is incorporated herein by reference. Asimilar technique, Nucleofection™, utilizes an applied electric field inorder to stimulate the uptake of exogenous polynucleotides into thenucleus of a eukaryotic cell. Nucleofection and protocols useful forperforming this technique are described in detail, e.g., in Distler etal. (2005) Experimental Dermatology 14:315, as well as in US2010/0317114, the disclosures of each of which are incorporated hereinby reference.

Additional techniques useful for the transfection of host cells for thepurposes of recombinant peptide and protein expression include thesqueeze-poration methodology. This technique induces the rapidmechanical deformation of cells in order to stimulate the uptake ofexogenous DNA through membranous pores that form in response to theapplied stress. This technology is advantageous in that a vector is notrequired for delivery of nucleic acids into a cell, such as a humancell. Squeeze-poration is described in detail, e.g., in Sharei et al.(2013) Journal of Visualized Experiments 81:e50980, the disclosure ofwhich is incorporated herein by reference.

Lipofection represents another technique useful for transfection ofcells. This method involves the loading of nucleic acids into aliposome, which often presents cationic functional groups, such asquaternary or protonated amines, towards the liposome exterior. Thispromotes electrostatic interactions between the liposome and a cell dueto the anionic nature of the cell membrane, which ultimately leads touptake of the exogenous nucleic acids, for example, by direct fusion ofthe liposome with the cell membrane or by endocytosis of the complex.Lipofection is described in detail, for example, in U.S. Pat. No.7,442,386, the disclosure of which is incorporated herein by reference.Similar techniques that exploit ionic interactions with the cellmembrane to provoke the uptake of foreign nucleic acids includecontacting a cell with a cationic polymer-nucleic acid complex.Exemplary cationic molecules that associate with polynucleotides so asto impart a positive charge favorable for interaction with the cellmembrane are activated dendrimers (described, e.g., in Dennig (2003)Topics in Current Chemistry 228:227, the disclosure of which isincorporated herein by reference) and diethylaminoethyl (DEAE)-dextran,the use of which as a transfection agent is described in detail, forexample, in Gulick et al. (1997) Current Protocols in Molecular Biology40:1:9.2:9.2.1, the disclosure of which is incorporated herein byreference. Magnetic beads are another tool that can be used to transfectcells in a mild and efficient manner, as this methodology utilizes anapplied magnetic field in order to direct the uptake of nucleic acids.This technology is described in detail, for example, in US 2010/0227406,the disclosure of which is incorporated herein by reference.

Another useful tool for inducing the uptake of exogenous nucleic acidsby cells is laserfection, a technique that involves exposing a cell toelectromagnetic radiation of a particular wavelength in order to gentlypermeabilize the cells and allow polynucleotides to penetrate the cellmembrane. This technique is described in detail, e.g., in Rhodes et al.(2007) Methods in Cell Biology 82:309, the disclosure of which isincorporated herein by reference.

Microvesicles represent another potential vehicle that can be used tointroduce a nucleic acid encoding a peptide or protein described hereininto a host cell for the purpose of recombinant expression. In someembodiments, microvesicles that have been induced by theco-overexpression of the glycoprotein VSV-G with, e.g., agenome-modifying protein, such as a nuclease, can be used to efficientlydeliver proteins into a cell that subsequently catalyze thesite-specific cleavage of an endogenous polynucleotide sequence so as toprepare the genome of the cell for the covalent incorporation of apolynucleotide of interest, such as a gene or regulatory sequence. Theuse of such vesicles, also referred to as Gesicles, for the geneticmodification of eukaryotic cells is described in detail, e.g., in Quinnet al., Genetic Modification of Target Cells by Direct Delivery ofActive Protein [abstract]. In: Methylation changes in early embryonicgenes in cancer [abstract], in: Proceedings of the 18th Annual Meetingof the American Society of Gene and Cell Therapy; 2015 May 13, AbstractNo. 122.

Viral Vectors for Nucleic Acid Delivery

Viral genomes provide a rich source of vectors that can be used for theefficient delivery of exogenous nucleic acids encoding peptides andproteins described herein, such as CXCR2 agonists, including Gro-β,Gro-β T, Gro-β N69D, Gro-β T N65D, and variants thereof, into host cellsfor the purpose of recombinant expression. Viral genomes areparticularly useful vectors for gene delivery because thepolynucleotides contained within such genomes may be incorporated intothe genome of a cell, for example, by way of generalized or specializedtransduction. These processes may occur as part of the naturalreplication cycle of a viral vector, and may not require added proteinsor reagents in order to induce gene integration. Examples of viralvectors that may be used to introduce a nucleic acid molecule encoding apeptide or protein described herein into a host cell for recombinantexpression include parvovirus, such as adeno-associated virus (AAV),retrovirus, adenovirus (e.g., Ad5, Ad26, Ad34, Ad35, and Ad48),coronavirus, negative strand RNA viruses such as orthomyxovirus (e.g.,influenza virus), rhabdovirus (e.g., rabies and vesicular stomatitisvirus), paramyxovirus (e.g. measles and Sendai), positive strand RNAviruses, such as picornavirus and alphavirus, and double stranded DNAviruses including adenovirus, herpesvirus (e.g., Herpes Simplex virustypes 1 and 2, Epstein-Barr virus, cytomegalovirus), and poxvirus (e.g.,vaccinia, modified vaccinia Ankara (MVA), fowlpox and canarypox). Otherviruses useful for delivering polynucleotides encoding peptides andproteins described herein to host cells for recombinant expressionpurposes include Norwalk virus, togavirus, flavivirus, reoviruses,papovavirus, hepadnavirus, and hepatitis virus, for example. Examples ofretroviruses include avian leukosis-sarcoma, mammalian C-type, B-typeviruses, D-type viruses, HTLV-BLV group, lentivirus, spumavirus (Coffin,J. M., Retroviridae: The viruses and their replication, In FundamentalVirology, Third Edition, B. N. Fields, et al., Eds., Lippincott-RavenPublishers, Philadelphia, 1996). Other examples include murine leukemiaviruses, murine sarcoma viruses, mouse mammary tumor virus, bovineleukemia virus, feline leukemia virus, feline sarcoma virus, avianleukemia virus, human T-cell leukemia virus, baboon endogenous virus,Gibbon ape leukemia virus, Mason Pfizer monkey virus, simianimmunodeficiency virus, simian sarcoma virus, Rous sarcoma virus andlentiviruses. Other examples of vectors are described, for example, inU.S. Pat. No. 5,801,030, the disclosure of which is incorporated hereinby reference as it pertains to viral vectors for use in gene deliveryand recombinant protein and peptide expression.

Methods of Treatment

As described herein, hematopoietic stem cell transplant therapy can beadministered to a subject in need of treatment so as to populate orrepopulate one or more blood cell types, such as a blood cell lineagethat is deficient or defective in a patient suffering from a stem celldisorder. Hematopoietic stem and progenitor cells exhibit multi-potency,and can thus differentiate into multiple different blood lineagesincluding, but not limited to, granulocytes (e.g., promyelocytes,neutrophils, eosinophils, basophils), erythrocytes (e.g., reticulocytes,erythrocytes), thrombocytes (e.g., megakaryoblasts, platelet producingmegakaryocytes, platelets), monocytes (e.g., monocytes, macrophages),dendritic cells, microglia, osteoclasts, and lymphocytes (e.g., NKcells, B-cells and T-cells). Hematopoietic stem cells are additionallycapable of self-renewal, and can thus give rise to daughter cells thathave equivalent potential as the mother cell, and also feature thecapacity to be reintroduced into a transplant recipient whereupon theyhome to the hematopoietic stem cell niche and re-establish productiveand sustained hematopoiesis. Thus, hematopoietic stem and progenitorcells represent a useful therapeutic modality for the treatment of awide array of disorders in which a patient has a deficiency or defect ina cell type of the hematopoietic lineage. The deficiency or defect maybe caused, for example, by depletion of a population of endogenous cellsof the hematopoietic system due to administration of a chemotherapeuticagent (e.g., in the case of a patient suffering from a cancer, such as ahematologic cancer described herein). The deficiency or defect may becaused, for example, by depletion of a population of endogenoushematopoietic cells due to the activity of self-reactive immune cells,such as T lymphocytes or B lymphocytes that cross-react with selfantigens (e.g., in the case of a patient suffering from an autoimmunedisorder, such as an autoimmune disorder described herein). Additionallyor alternatively, the deficiency or defect in cellular activity may becaused by aberrant expression of an enzyme (e.g., in the case of apatient suffering from various metabolic disorders, such as a metabolicdisorder described herein).

Thus, hematopoietic stem cells can be administered to a patientdefective or deficient in one or more cell types of the hematopoieticlineage in order to re-constitute the defective or deficient populationof cells in vivo, thereby treating the pathology associated with thedefect or depletion in the endogenous blood cell population.Hematopoietic stem and progenitor cells can be used to treat, e.g., anon-malignant hemoglobinopathy (e.g., a hemoglobinopathy selected fromthe group consisting of sickle cell anemia, thalassemia, Fanconi anemia,aplastic anemia, and Wiskott-Aldrich syndrome). In these cases, forexample, a CXCR4 antagonist and/or a CXCR2 agonist may be administeredto a donor, such as a donor identified as likely to exhibit release of apopulation of hematopoietic stem and progenitor cells from a stem cellniche, such as the bone marrow, into circulating peripheral blood inresponse to such treatment. The hematopoietic stem and progenitor cellsthus mobilized may then be withdrawn from the donor and administered toa patient, where the cells may home to a hematopoietic stem cell nicheand re-constitute a population of cells that are damaged or deficient inthe patient.

Additionally or alternatively, hematopoietic stem and progenitor cellscan be used to treat an immunodeficiency, such as a congenitalimmunodeficiency. Additionally or alternatively, the compositions andmethods described herein can be used to treat an acquiredimmunodeficiency (e.g., an acquired immunodeficiency selected from thegroup consisting of HIV and AIDS). In these cases, for example, a CXCR4antagonist and/or a CXCR2 agonist may be administered to a donor, suchas a donor identified as likely to exhibit release of a population ofhematopoietic stem and progenitor cells from a stem cell niche, such asthe bone marrow, into circulating peripheral blood in response to suchtreatment. The hematopoietic stem and progenitor cells thus mobilizedmay then be withdrawn from the donor and administered to a patient,where the cells may home to a hematopoietic stem cell niche andre-constitute a population of immune cells (e.g., T lymphocytes, Blymphocytes, NK cells, or other immune cells) that are damaged ordeficient in the patient.

Hematopoietic stem and progenitor cells can also be used to treat ametabolic disorder (e.g., a metabolic disorder selected from the groupconsisting of glycogen storage diseases, mucopolysaccharidoses,Gaucher's Disease, Hurlers Disease, sphingolipidoses, and metachromaticleukodystrophy). In these cases, for example, a CXCR4 antagonist and/ora CXCR2 agonist may be administered to a donor, such as a donoridentified as likely to exhibit release of a population of hematopoieticstem and progenitor cells from a stem cell niche, such as the bonemarrow, into circulating peripheral blood in response to such treatment.The hematopoietic stem and progenitor cells thus mobilized may then bewithdrawn from the donor and administered to a patient, where the cellsmay home to a hematopoietic stem cell niche and re-constitute apopulation of hematopoietic cells that are damaged or deficient in thepatient.

Additionally or alternatively, hematopoietic stem or progenitor cellscan be used to treat a malignancy or proliferative disorder, such as ahematologic cancer or myeloproliferative disease. In the case of cancertreatment, for example, a CXCR4 antagonist and/or a CXCR2 agonist may beadministered to a donor, such as a donor identified as likely to exhibitrelease of a population of hematopoietic stem and progenitor cells froma stem cell niche, such as the bone marrow, into circulating peripheralblood in response to such treatment. The hematopoietic stem andprogenitor cells thus mobilized may then be withdrawn from the donor andadministered to a patient, where the cells may home to a hematopoieticstem cell niche and re-constitute a population of cells that are damagedor deficient in the patient, such as a population of hematopoietic cellsthat is damaged or deficient due to the administration of one or morechemotherapeutic agents to the patient. In some embodiments,hematopoietic stem or progenitor cells may be infused into a patient inorder to repopulate a population of cells depleted during cancer celleradication, such as during systemic chemotherapy. Exemplaryhematological cancers that can be treated by way of administration ofhematopoietic stem and progenitor cells in accordance with thecompositions and methods described herein are acute myeloid leukemia,acute lymphoid leukemia, chronic myeloid leukemia, chronic lymphoidleukemia, multiple myeloma, diffuse large B-cell lymphoma, andnon-Hodgkin's lymphoma, as well as other cancerous conditions, includingneuroblastoma.

Hematopoietic stem or progenitor cells mobilized to the peripheral bloodof a subject may be withdrawn (e.g., harvested or collected) from thesubject by any suitable technique. For example, the hematopoietic stemor progenitor cells may be withdrawn by a blood draw. In someembodiments, hematopoietic stem or progenitor cells mobilized to asubject's peripheral blood as contemplated herein may be harvested(i.e., collected) using apheresis. In some embodiments, apheresis may beused to enrich a donor's blood with mobilized hematopoietic stem orprogenitor cells.

Additional diseases that can be treated by the administration ofhematopoietic stem and progenitor cells to a patient include, withoutlimitation, adenosine deaminase deficiency and severe combinedimmunodeficiency, hyper immunoglobulin M syndrome, Chediak-Higashidisease, hereditary lymphohistiocytosis, osteopetrosis, osteogenesisimperfecta, storage diseases, thalassemia major, systemic sclerosis,systemic lupus erythematosus, multiple sclerosis, and juvenilerheumatoid arthritis.

In addition, administration of hematopoietic stem and progenitor cellscan be used to treat autoimmune disorders. In some embodiments, uponinfusion into a patient, transplanted hematopoietic stem and progenitorcells may home to a stem cell niche, such as the bone marrow, andestablish productive hematopoiesis. This, in turn, can re-constitute apopulation of cells depleted during autoimmune cell eradication, whichmay occur due to the activity of self-reactive lymphocytes (e.g.,self-reactive T lymphocytes and/or self-reactive B lymphocytes).Autoimmune diseases that can be treated by way of administeringhematopoietic stem and progenitor cells to a patient include, withoutlimitation, psoriasis, psoriatic arthritis, Type 1 diabetes mellitus(Type 1 diabetes), rheumatoid arthritis (RA), human systemic lupus(SLE), multiple sclerosis (MS), inflammatory bowel disease (IBD),lymphocytic colitis, acute disseminated encephalomyelitis (ADEM),Addison's disease, alopecia universalis, ankylosing spondylitis,antiphospholipid antibody syndrome (APS), aplastic anemia, autoimmunehemolytic anemia, autoimmune hepatitis, autoimmune inner ear disease(AIED), autoimmune lymphoproliferative syndrome (ALPS), autoimmuneoophoritis, Balo disease, Behcet's disease, bullous pemphigoid,cardiomyopathy, Chagas' disease, chronic fatigue immune dysfunctionsyndrome (CFIDS), chronic inflammatory demyelinating polyneuropathy,Crohn's disease, cicatricial pemphigoid, coeliac sprue-dermatitisherpetiformis, cold agglutinin disease, CREST syndrome, Degos disease,discoid lupus, dysautonomia, endometriosis, essential mixedcryoglobulinemia, fibromyalgia-fibromyositis, Goodpasture's syndrome,Grave's disease, Guillain-Barre syndrome (GBS), Hashimoto's thyroiditis,Hidradenitis suppurativa, idiopathic and/or acute thrombocytopenicpurpura, idiopathic pulmonary fibrosis, IgA neuropathy, interstitialcystitis, juvenile arthritis, Kawasaki's disease, lichen planus, Lymedisease, Meniere disease, mixed connective tissue disease (MCTD),myasthenia gravis, neuromyotonia, opsoclonus myoclonus syndrome (OMS),optic neuritis, Ord's thyroiditis, pemphigus vulgaris, perniciousanemia, polychondritis, polymyositis and dermatomyositis, primarybiliary cirrhosis, polyarteritis nodosa, polyglandular syndromes,polymyalgia rheumatica, primary agammaglobulinemia, Raynaud phenomenon,Reiter's syndrome, rheumatic fever, sarcoidosis, scleroderma, Sjögren'ssyndrome, stiff person syndrome, Takayasu's arteritis, temporalarteritis (also known as “giant cell arteritis”), ulcerative colitis,collagenous colitis, uveitis, vasculitis, vitiligo, vulvodynia (“vulvarvestibulitis”), and Wegener's granulomatosis.

In some embodiments, a method of harvesting hematopoietic stem cellsfrom a human subject is provided. The method comprises administering aCXCR2 agonist and a CXCR4 antagonist to the human subject and harvestingthe hematopoietic stem cells from peripheral blood of the human subject.

In some embodiments, a method of transplanting hematopoietic stem cellsinto a human patient in need thereof is provided. The method comprisesadministering a CXCR2 agonist and a CXCR4 antagonist to a hematopoieticstem cell donor, harvesting the hematopoietic stem cells from peripheralblood of the donor, and transplanting the harvested hematopoietic stemcells into the patient.

Selection of Donors and Patients

In some embodiments, the patient is the donor. In such cases, withdrawnhematopoietic stem or progenitor cells may be re-infused into thepatient, such that the cells may subsequently home hematopoietic tissueand establish productive hematopoiesis, thereby populating orrepopulating a line of cells that is defective or deficient in thepatient (e.g., a population of megakaryocytes, thrombocytes, platelets,erythrocytes, mast cells, myeoblasts, basophils, neutrophils,eosinophils, microglia, granulocytes, monocytes, osteoclasts,antigen-presenting cells, macrophages, dendritic cells, natural killercells, T-lymphocytes, and B-lymphocytes). In this scenario, thetransplanted hematopoietic stem or progenitor cells are least likely toundergo graft rejection, as the infused cells are derived from thepatient and express the same HLA class I and class II antigens asexpressed by the patient.

Alternatively, the patient and the donor may be distinct. In someembodiments, the patient and the donor are related, and may, forexample, be HLA-matched. As described herein, HLA-matcheddonor-recipient pairs have a decreased risk of graft rejection, asendogenous T cells and NK cells within the transplant recipient are lesslikely to recognize the incoming hematopoietic stem or progenitor cellgraft as foreign, and are thus less likely to mount an immune responseagainst the transplant. Exemplary HLA-matched donor-recipient pairs aredonors and recipients that are genetically related, such as familialdonor-recipient pairs (e.g., sibling donor-recipient pairs).

In some embodiments, the patient and the donor are HLA-mismatched, whichoccurs when at least one HLA antigen, in particular with respect toHLA-A, HLA-B and HLA-DR, is mismatched between the donor and recipient.To reduce the likelihood of graft rejection, for example, one haplotypemay be matched between the donor and recipient, and the other may bemismatched.

CD34^(dim) Cells

When the donor and patient are distinct, methods of treatment usinghematopoietic stem cells comprising CD34^(dim) cells are particularlyuseful, in part because CD34^(dim) cells are capable of suppressingalloreactive T lymphocyte proliferation when administered to arecipient, thereby reducing the risk of graft vs. host disease (GVHD).(D'Aveni et al. (2015), supra.) In certain embodiments, administrationof a CXCR2 agonist and a CXCR4 antagonist according to the methodsdisclosed herein mobilizes hematopoietic stem cells comprisingCD34^(dim) cells from the bone marrow of a donor into peripheral blood.The CD34^(dim) cells are present in a higher amount in the peripheralblood as compared to peripheral blood from an unmobilized mammal. Incertain embodiments, the CD34^(dim) cells are present in a higher amountin the peripheral blood than if the hematopoietic stem cells weremobilized using the CXCR4 antagonist alone.

Accordingly, the methods disclosed herein are useful in performing anallogeneic hematopoietic stem cell transplant in a patient in needthereof. For example, the method can include infusing into the patient atherapeutically effective amount of allogeneic hematopoietic stem cells,wherein the hematopoietic stem cells were mobilized from bone marrow ofa human donor into peripheral blood of the human donor using the methodsherein. In certain embodiments, the method includes administering to thedonor (i) a CXCR2 agonist selected from the group consisting of Gro-β,Gro-β T, and variants thereof at a dose of from about 50 μg/kg to about1,000 μg/kg and (ii) a CXCR4 antagonist.

In addition, CD34^(dim) cells have been shown to increase overallsurvival (OS), decrease non-relapse mortality (NRM, i.e., the time todeath without relapse/recurrence), and lower the risk of infection(e.g., cytomegalovirus (CMV) infection) in a patient having a standardrisk disease receiving an allogenic hematopoietic cell transplant.(Nakasone et al. “CD34⁺ monocytes mobilized by G-CSF in donor PB andclinical outcomes after all-HCT from related donors,” Poster presentedat 44^(th) Annual Meeting of the European Society for Blood and MarrowTransplantation, Mar. 18-21, 2018, Lisbon, Portugal.)

Accordingly, in certain embodiments, the methods of treating a stem celldisorder in a human patient disclosed herein can include infusing intothe patient a therapeutically effective amount of the hematopoietic stemcells mobilized by any one of the methods disclosed herein, wherein themobilized hematopoietic stem cells comprise CD34^(dim) cells, andwherein the treatment results in increased OS, decreased NRM, and/orlowered risk of infection (e.g., CMV infection).

In addition, the methods described herein can be used in preventing,reducing the risk of developing, or reducing the severity of apost-transplant infection in a patient in need thereof. The method caninclude infusing into the patient a therapeutically effective amount ofhematopoietic stem cells, wherein the hematopoietic stem cells weremobilized from bone marrow of a human donor into peripheral blood of thehuman donor according to the methods described herein, for example,administering to the human donor (i) a CXCR2 agonist selected from thegroup consisting of Gro-β, Gro-β T, and variants thereof at a dose offrom about 50 μg/kg to about 1,000 μg/kg and (ii) a CXCR4 antagonist. Incertain embodiments, the infection is a CMV infection.

In addition, the disclosure relates to a method of preventing, reducingthe risk of developing, or reducing the severity of graft versus hostdisease (GVHD) in a patient in need thereof, wherein the method includesinfusing into the patient a therapeutically effective amount ofhematopoietic stem cells, wherein the hematopoietic stem cells weremobilized from bone marrow of a mammalian donor into peripheral blood bythe methods described herein, e.g., including administering to themammalian donor a CXCR2 agonist and a CXCR4 antagonist.

In certain embodiments, the hematopoietic stem and progenitor cellsmobilized from the bone marrow of a donor into peripheral blood compriseat least 1%, at least 2% at least 5% at least 10%, at least 15%, atleast 20% or at least 20% or more CD34^(dim) cells as compared toperipheral blood from an unmobilized mammal. In certain embodiments, thehematopoietic stem and progenitor cells mobilized from the bone marrowof a donor into peripheral blood comprise at from about 1% to about 5%,from about 1% to about 10%, from about 1% to about 15%, from about 1% toabout 20%, from about 2% to about 25%, 2% to about 5%, from about 2% toabout 10%, from about 2% to about 15%, from about 2% to about 20%, fromabout 2% to about 25%, from about 5% to about 10%, from about 5% toabout 15%, from about 5% to about 20%, from about 5% to about 25%, fromabout 10% to about 15%, from about 10% to about 20%, from about 10% toabout 25%, from about 15% to about 20%, from about 15% to about 25%CD34^(dim) cells as compared to peripheral blood from an immobilizedmammal.

In certain embodiments, the hematopoietic stem and progenitor cellsmobilized from the bone marrow of a donor into peripheral blood compriseat least 1.5-fold, at least 2-fold, at least 3-fold, at least 4-fold, atleast 5-fold, at least 6-fold, at least 7-fold, at least 8-fold, atleast 9-fold, at least 10-fold, at least 11-fold, at least 12-fold, atleast 13-fold, at least 14-fold, at least 15-fold, at least 20-fold, atleast 30-fold, at least 50-fold more CD34^(dim) cells as compared toperipheral blood from an unmobilized mammal. In certain embodiments, thehematopoietic stem and progenitor cells mobilized from the bone marrowof a donor into peripheral blood comprise between about 1.5-fold and30-fold, between about 5-fold and about 25-fold, between about 10-foldand about 20-fold, or between about 12-fold and about 17-fold moreCD34^(dim) cells as compared to peripheral blood from an unmobilizedmammal.

In certain embodiments, the hematopoietic stem and progenitor cellsmobilized from the bone marrow of a donor into peripheral blood compriseat least 1%, at least 2% at least 5% at least 10%, at least 15%, atleast 20% or at least 20% or more CD34^(dim) cells than if thehematopoietic stem cells were mobilized using the CXCR4 antagonistalone. In certain embodiments, the hematopoietic stem and progenitorcells mobilized from the bone marrow of a donor into peripheral bloodcomprise at from about 1% to about 5%, from about 1% to about 10%, fromabout 1% to about 15%, from about 1% to about 20%, from about 2% toabout 25%, 2% to about 5%, from about 2% to about 10%, from about 2% toabout 15%, from about 2% to about 20%, from about 2% to about 25%, fromabout 5% to about 10%, from about 5% to about 15%, from about 5% toabout 20%, from about 5% to about 25%, from about 10% to about 15%, fromabout 10% to about 20%, from about 10% to about 25%, from about 15% toabout 20%, from about 15% to about 25% CD34^(dim) cells than if thehematopoietic stem cells were mobilized using the CXCR4 antagonistalone.

In certain embodiments, the hematopoietic stem and progenitor cellsmobilized from the bone marrow of a donor into peripheral blood compriseat least 1.5-fold, at least 2-fold, at least 3-fold, at least 4-fold, atleast 5-fold, at least 6-fold, at least 7-fold, at least 8-fold, atleast 9-fold, at least 10-fold, at least 11-fold, at least 12-fold, atleast 13-fold, at least 14-fold, at least 15-fold, at least 20-fold, atleast 30-fold, at least 50-fold more CD34^(dim) cells than if thehematopoietic stem cells were mobilized using the CXCR4 antagonistalone. In certain embodiments, the hematopoietic stem and progenitorcells mobilized from the bone marrow of a donor into peripheral bloodcomprise between about 1.5-fold and 30-fold, between about 5-fold andabout 25-fold, between about 10-fold and about 20-fold, or between about12-fold and about 17-fold more CD34^(dim) cells than if thehematopoietic stem cells were mobilized using the CXCR4 antagonistalone.

Methods of Genetic Modification of Hematopoietic Stem and ProgenitorCells

Prior to infusion into a patient, such as a patient having one or morestem cell disorders described herein, hematopoietic stem cells obtainedfrom a donor (or progeny thereof) may be genetically modified, forexample, by disrupting an endogenous gene. This strategy can be used,for example, to silence the expression of one or more majorhistocompatibility complex genes in a hematopoietic stem cell that isallogeneic with respect to the patient, thereby reducing the likelihoodof graft rejection upon transplantation.

A wide array of methods has been established for the disruption oftarget genes in a population of cells. In some embodiments, one suchmethod is through the use of a clustered regularly interspaced shortpalindromic repeats (CRISPR)/Cas system, a system that originallyevolved as an adaptive defense mechanism in bacteria and archaea againstviral infection. The CRISPR/Cas system includes palindromic repeatsequences within plasmid DNA and an associated Cas9 nuclease. Thisensemble of DNA and protein directs site specific DNA cleavage of atarget sequence by first incorporating foreign DNA into CRISPR loci.Polynucleotides containing these foreign sequences and the repeat-spacerelements of the CRISPR locus are in turn transcribed in a host cell tocreate a guide RNA, which can subsequently anneal to a target sequenceand localize the Cas9 nuclease to this site. In this manner, highlysite-specific cas9-mediated DNA cleavage can be engendered in a foreignpolynucleotide because the interaction that brings cas9 within closeproximity of the target DNA molecule is governed by RNA:DNAhybridization. As a result, one can theoretically design a CRISPR/Cassystem to cleave any target DNA molecule of interest. This technique hasbeen exploited in order to edit eukaryotic genomes (Hwang et al. (2013)Nature Biotechnology 31:227, the disclosure of which is incorporatedherein by reference) and can be used as an efficient means ofsite-specifically editing hematopoietic stem cell genomes in order tocleave DNA, for example, prior to the incorporation of a gene encoding atarget protein. The use of CRISPR/Cas to modulate gene expression hasbeen described in, e.g., U.S. Pat. No. 8,697,359, the disclosure ofwhich is incorporated herein by reference. Alternative methods forsite-specifically cleaving genomic DNA prior to the incorporation of agene of interest in a hematopoietic stem cell include the use of zincfinger nucleases (ZFNs) and transcription activator-like effectornucleases (TALENs). Unlike the CRISPR/Cas system, these enzymes do notcontain a guiding polynucleotide to localize to a specific targetsequence. Target specificity is instead controlled by DNA bindingdomains within these enzymes. The use of ZFNs and TALENs in genomeediting applications is described, e.g., in Urnov et al. (2010) NatureReviews Genetics 11:636; and in Joung et al. (2013) Nature ReviewsMolecular Cell Biology 14:49, the disclosure of both of which areincorporated herein by reference.

Additional genome editing techniques that can be used to incorporatepolynucleotides encoding target genes into the genome of a hematopoieticstem cell include the use of ARCUS™ meganucleases that can be rationallydesigned so as to site-specifically cleave genomic DNA. The use of theseenzymes for the incorporation of genes encoding target genes into thegenome of a mammalian cell is advantageous in view of the definedstructure-activity relationships that have been established for suchenzymes. Single chain meganucleases can be modified at certain aminoacid positions in order to create nucleases that selectively cleave DNAat desired locations, enabling the site-specific incorporation of atarget gene into the nuclear DNA of a hematopoietic stem cell. Thesesingle-chain nucleases have been described extensively in, e.g., U.S.Pat. Nos. 8,021,867 and 8,445,251, the disclosures of each of which areincorporated herein by reference.

Kinetics of CXCR2 Agonist and CXCR4 Antagonist Dosing

For cases in which the donor is administered both a CXCR4 antagonist anda CXCR2 agonist, the two agents may be administered to the donorconcurrently. In some embodiments, the CXCR4 antagonist and the CXCR2agonist may be co-formulated with one another and administered in thesame pharmaceutical composition. Alternatively, the CXCR4 antagonist andthe CXCR2 agonist may be formulated in distinct pharmaceuticalcompositions and administered separately but simultaneously to thedonor.

In some embodiments, the CXCR4 antagonist is administered to the donorprior to administration of the CXCR2 agonist. In some embodiments, theCXCR4 antagonist may be administered to the donor from about 30 minutesto about 180 minutes prior to administration of the CXCR2 agonist, suchas from about 40 minutes to about 160 minutes, about 50 minutes to about150 minutes, about 60 minutes to about 140 minutes, about 70 minutes toabout 130 minutes, about 60 minutes to about 120 minutes, about 70minutes to about 110 minutes, or about 80 minutes to about 100 minutes(e.g., about 30 minutes, about 35 minutes, about 40 minutes, about 45minutes, about 50 minutes, about 55 minutes, about 60 minutes, about 65minutes, about 70 minutes, about 75 minutes, about 80 minutes, about 85minutes, about 90 minutes, about 95 minutes, about 100 minutes, about105 minutes, about 110 minutes, about 115 minutes, about 120 minutes,about 125 minutes, about 130 minutes, about 135 minutes, about 140minutes, about 145 minutes, about 150 minutes, about 155 minutes, about160 minutes, about 165 minutes, about 170 minutes, about 175 minutes, orabout 180 minutes prior to administration of the CXCR2 agonist). In someembodiments, the CXCR4 antagonist is administered to the donor fromabout 30 minutes to about 60 minutes prior to administration of theCXCR2 agonist (e.g., about 30 minutes, about 35 minutes, about 40minutes, about 45 minutes, about 50 minutes, about 55 minutes, or about60 minutes prior to administration of the CXCR2 agonist). In someembodiments, the CXCR4 antagonist may be administered to the donor about45 minutes prior to administration of the CXCR2 agonist.

Isolation of the population of hematopoietic stem or progenitor cellsmay commence from about 10 minutes to about 60 minutes followingcompletion of the administration of the CXCR4 antagonist and the CXCR2agonist (e.g., about 10 minutes to about 1.9 hours, about 20 minutes toabout 1.8 hours, about 25 minutes to about 1.7 hours, about 30 minutesto about 1.6 hours, about 40 minutes to about 1.5 hours (e.g., about 10minutes, about 15 minutes, about 20 minutes, about 25 minutes, about 30minutes, about 35 minutes, about 40 minutes, about 45 minutes, about 50minutes, about 55 minutes, about 60 minutes, or about 120 minutesfollowing completion of the administration of the CXCR4 antagonist andthe CXCR2 agonist). In some embodiments, isolation of the population ofhematopoietic stem or progenitor cells may commence from about 10minutes to about 20 minutes following completion of the administrationof the CXCR4 antagonist and the CXCR2 agonist (e.g., about 10 minutes,about 11 minutes, about 12 minutes, about 13 minutes, about 14 minutes,about 15 minutes, about 16 minutes, about 17 minutes, about 18 minutes,about 19 minutes, or about 20 minutes following completion of theadministration of the CXCR4 antagonist and the CXCR2 agonist). In someembodiments, isolation of the population of hematopoietic stem orprogenitor cells commences about 15 minutes following completion of theadministration of the CXCR4 antagonist and the CXCR2 agonist.

In some embodiments, the population of hematopoietic stem or progenitorcells is isolated from the donor over a period of from about 15 minutesto about 6 hours, such as from about 20 minutes to about 4.5 hours,about 30 minutes to about 4 hours, about 40 minutes to about 3.5 hours,about 50 minutes to about 3 hours, or about 1 hour to about 2 hours(e.g., over a period of about 15 minutes, about 20 minutes, about 30minutes, about 35 minutes, about 40 minutes, about 45 minutes, about 50minutes, about 55 minutes, about 60 minutes, about 65 minutes, about 70minutes, about 75 minutes, about 80 minutes, about 85 minutes, about 90minutes, about 95 minutes, about 100 minutes, about 105 minutes, about110 minutes, about 115 minutes, about 120 minutes, about 180 minutes,about 240 minutes, about 300 minutes, or about 360 minutes). In someembodiments, the population of hematopoietic stem and progenitor cellsmay be isolated from the donor over a period of from about 30 minutes toabout 1 hour (e.g., over a period of about 30 minutes, about 35 minutes,about 40 minutes, about 45 minutes, about 50 minutes, about 55 minutes,or about 60 minutes).

In some embodiments, the hematopoietic stem or progenitor cells may beharvested by apheresis. In some embodiments, the hematopoietic stem orprogenitor cells may be harvested by drawing peripheral blood from thedonor (i.e., subject).

Routes of Administration of CXCR2 Agonists and CXCR4 Antagonists

The CXCR4 antagonists and CXCR2 agonists described herein may beadministered to a patient by a variety of routes, such as intravenously,subcutaneously, intramuscularly, or parenterally. The most suitableroute for administration in any given case will depend on the particularagent administered, the patient, pharmaceutical formulation methods,administration methods (e.g., administration time and administrationroute), the patient's age, body weight, sex, severity of the diseasesbeing treated, the patient's diet, and the patient's excretion rate.Preferably, the CXCR2 agonist (e.g., Gro-β, Gro-β T, or a variantthereof) may be administered to a donor intravenously. Under theseconditions, CXCR2 agonists, such as those described herein, rapidly giverise to populations of cells that are enriched in CD34⁺ CD90⁺ CD45RA⁻cells (hematopoietic stem cells), and reduce the mobilization of othercell types, such as leukocytes, neutrophils, lymphocytes, and monocytes.This property is described in further detail in Example 1, below.

Pharmaceutical Compositions

The CXCR2 agonists and CXCR4 antagonists contemplated herein may each beformulated into a pharmaceutical composition for administration to asubject, such as a mammalian subject (e.g., a human subject). Forinstance, contemplated herein are pharmaceutical compositions comprisinga CXCR2 agonist and/or a CXCR4 antagonist, in admixture with one or moresuitable diluents, carriers, and/or excipients. Pharmaceuticalcompositions may include sterile aqueous suspensions. Conventionalprocedures and ingredients for the selection and preparation of suitableformulations are described, for example, in Remington: The Science andPractice of Pharmacy (2012, 22^(nd) ed.) and in The United StatesPharmacopeia: The National Formulary (2015, USP 38 NF 33), thedisclosure of which is incorporated herein by reference in its entirety.

A pharmaceutical composition may be administered to a subject, such as ahuman subject, alone or in combination with pharmaceutically acceptablecarriers, the proportion of which may be determined by the quantity ofactive pharmaceutical ingredient (i.e., CXCR2 agonist and/or a CXCR4antagonist), chosen route of administration, and standard pharmaceuticalpractice.

Administration and Dosing of CXCR2 Agonists and/or CXCR4 Antagonists

Contemplated CXCR2 agonists and CXCR4 antagonists, may be administeredto a subject, such as a mammalian subject (e.g., a human subject), byone or more routes of administration. For instance, contemplated CXCR2agonists and CXCR4 antagonists may be administered to a subject byintravenous, intraperitoneal, intramuscular, intraarterial, orsubcutaneous infusion, among others.

Contemplated CXCR2 agonists and CXCR4 antagonists may be administered toa subject in one or more doses. For example, a CXCR2 agonist and/orCXCR4 antagonist may be administered as a single dose or in two, three,four, five, or more doses. When multiple doses are administered,subsequent doses may be provided during the same day or one or moredays, weeks, months, or years following the initial dose. For instance,the contemplated CXCR2 agonists and CXCR4 antagonists described hereinmay be administered to a subject, such as a human subject one or moretimes daily, weekly, monthly, or yearly, depending on such factors as,for instance, the subject's age, body weight, sex, the subject's diet,and the subject's excretion rate. In certain embodiments, thecontemplated CXCR2 agonists and CXCR4 antagonists are each administeredin a single dose once per day.

Hematopoietic stem or progenitor cells and pharmaceutical compositionsdescribed herein may be administered to a subject in one or more doses.When multiple doses are administered, subsequent doses may be providedone or more days, weeks, months, or years following the initial dose.For instance, the hematopoietic stem cells and pharmaceuticalcompositions described herein may be administered to a subject, such asa human subject suffering from one or more diseases, conditions, ordisorders described herein, one or more times daily, weekly, monthly, oryearly, depending on such factors as, for instance, the subject's age,body weight, sex, severity of the diseases being treated, the subject'sdiet, and the subject's excretion rate.

EXAMPLES

The following examples are put forth so as to provide those of ordinaryskill in the art with a description of how the compositions and methodsdescribed herein may be used, made, and evaluated, and are intended tobe purely exemplary of the invention and are not intended to limit thescope of what the inventors regard as their invention.

Example 1. The Effects of Gro-β T on the Mobilization of HematopoieticStem Cells in Mice and Rhesus Monkeys

Mobilized peripheral blood grafts are currently the predominant sourceof hematopoietic stem and progenitor cells (HSPC) for both autologousand allogeneic transplantation. The most common clinical hematopoieticstem cell mobilization protocol is five days of Filgrastim (G-CSF). Thisregimen requires daily injections, has been associated with bone painand often results in unpredictably low yields. A rapid mobilizationmethod that ideally only required a single treatment and had robust andpredictable kinetics would be a significant improvement over the currentstandard of care. In mice, a unique CXCR2 agonist, T, induces rapidmobilization of stem and progenitor cells 15 minutes after a singleinjection. When co-administered with plerixafor (AMD3100), an inhibitorof CXCR4, a synergistic increase in mobilization results, and grafts areenriched in highly engraftable, long-term hematopoietic stem cells (see,e.g., FIG. 1A; LT-HSC=Lin-c-kit⁺ Sca-1⁺ CD150⁺ CD48⁺). Grafts containingcells mobilized by Gro-β T and plerixaflor led to higher relativenumbers of competitive repopulating units (CRU) at week 16 than didgrafts containing cells mobilized by G-CSF alone (FIG. 1B).

In this example, data are presented to demonstrate that combinationtreatment with Gro-β T and AMD3100 results in significantly enhancedmobilization of CD34⁺ cells and colony forming units (CFU) compared tothat achieved with AMD3100 alone in nonhuman primates (NHP).

Mobilization of hematopoietic stem cells was investigated in rhesusmacaques using Gro-β T and plerixafor (also referred to as AMD3100) asdescribed below.

Methods

Male rhesus macaques were treated with AMD3100 alone or in combinationwith Gro-β T. Blood was collected immediately prior to and 0.5, 1, 2, 4and 24 hours after treatment and analyzed by multicolor flow cytometryto quantitate HSPC numbers. Additional aliquots of mobilized blood wereplated in methylcellulose and CFU were enumerated seven days later.

Results

FIG. 2A shows the pharmacokinetic profile of various dosages of Gro-β Twhen administered intravenously to Rhesus monkeys. FIG. 2B shows thepharmacokinetic profile of various dosages of Gro-β T when administeredsubcutaneously to Rhesus monkeys. In all experiments, Gro-β T wasadministered to subjects concurrently with plerixafor.

As shown in FIGS. 3-6, Gro-β T and AMD3100 mobilizes white blood cellsinto the peripheral blood. Animals were mobilized with AMD3100 alone orin combination with Gro-β T. Peripheral blood was collected at the timepoints shown and white blood cells enumerated on a HESKA HematologyAnalyzer. Total number of white blood cells, neutrophils, lymphocytesand monocytes per μL of peripheral blood were determined. Data shown inFIGS. 2-5 are expressed as mean±SEM and represent 5 animals per group.Statistical significance was determined based on 2-way ANOVA withpost-hoc Dunnett's multiple comparisons test (** p<0.01).

Importantly, as shown in FIGS. 7 and 8, Gro-β T and AMD3100 inducesrobust mobilization of CD34⁺ cells into the peripheral blood. The datashown in FIGS. 7 and 8 include the percentage of whole blood accountedfor by CD34⁺ hematopoietic stem and progenitor cells for each treatmentgroup. Absolute numbers and fold change in CD34⁺ cells per μL ofperipheral blood were quantified with a single platform quantitativemethod. Data shown in FIGS. 7 and 8 are expressed as mean±SEM andrepresent 5 animals per group. Statistical significance was determinedbased on 2-way ANOVA with post-hoc Dunnett's multiple comparisons test(* p<0.05, ** p<0.01, **** p<0.0001).

Additionally, as shown in FIGS. 9 and 10, CD34⁺ cells mobilized inresponse to Gro-β T+AMD3100 are enriched for primitive CD34⁺CD90+CD45RA⁻ stem and progenitor cells. The data shown in FIGS. 9 and 10include the percentage of whole blood accounted for by CD34⁺ CD90⁺CD45RA⁻ hematopoietic stem and progenitor cells for each treatmentgroup. Absolute numbers and fold change in CD34⁺ CD90⁺ CD45RA⁻ cells perμL of peripheral blood are shown. Data shown in FIGS. 9 and 10 areexpressed as mean±SEM and represent 5 animals per group. Statisticalsignificance was determined based on 2-way ANOVA with post-hoc Dunnett'smultiple comparisons test (* p<0.05. **p<0.01, *** p<0.001, ****p<0.0001).

Further, as shown in FIG. 11, Gro-β T and AMD3100 mobilizeshematopoietic stem and progenitor cells with colony forming potential.The number of CFU per mL of peripheral blood was enumerated after sevendays of culture in methylcellulose. Data shown in FIG. 11 are expressedas mean±SEM and represent 3-5 animals per group. Statisticalsignificance was determined based on 2-way ANOVA with post-hoc Dunnett'smultiple comparisons test (* p<0.05). The ratio of MMP-9 to TIMP-1 isadditionally elevated following treatment with Gro-β T and AMD3100(FIGS. 12-14).

Additional data summarizing the mobilization of CD34⁺ cells (e.g., CD34⁺CD90⁺ CD45RA⁻ cells) in Rhesus monkeys using various doses of Gro-beta Tand AMD3100 are reported in TABLES 8-11, below. Quantities are reportedin TABLES 8-11 using the following notation: “Median value (Minimumvalue observed−maximum value observed).”

TABLE 8 Mobilization response observed in Rhesus monkeys uponadministration of plerixafor (1 mg/kg, subcutaneously) Ratio of Ratio offold quantity of increase of Quantity of CD34⁺ CD34⁺ cells observedRatio of CD90⁺ Fold increase Ratio of fold CD90⁺ 4 hours quantity ofCD45RA⁻ vs. baseline increase of CD45RA⁻ following sc CD34⁺ cells cellsto following sc CD34⁺ cells cells to other administration to other cellother cell administration to other cell cell Cell type of AMD3100:populations populations of AMD3100: populations populations CD34⁺ 41054— 0.352 9.1 — 1.4 (23012- (0.259- (7.8-16.5) (0.6-2.6) 58496) 0.551)CD34⁺ 14820 2.838 — 14.9 0.7 — CD90⁺ (6123- (1.816- (5.3-23.3) (0.4-1.7)CD54RA⁻ 17893) 3.864) WBCs 3.90E+07 0.0009 0.0003 2.3 2.3 3.7(3.16-4.97E+07) (0.0006- (0.0002- (1.6-4.6) (1.6-4.6) (1.0-7.9) 0.0012)0.0004) Neutrophils 2.10E+07 0.0015 0.0006 5.0 2.1 3.0 (1.61-3.31E+07)(0.0011- (0.0004- (3.0-14.0) (0.6-3.6) (0.4-3.2) 0.0021) 0.0007)Lymphocytes 1.35E+07 0.0025 0.0011 2.5 4.0 5.7 (1.12-1.62E+07) (0.0020-(0.0005- (1.3-3.1)  (2.9-8.2)  (2.4-14.0) 0.0043) 0.0013) Monocytes3.10E+06 0.0111 0.0039 7.9 1.2 1.6 (1.55-4.93E+06) (0.0047- (0.0020-(5.4-14.1) (0.6-2.0) (0.7-3.5) 0.0377) 0.0115)

TABLE 9 Mobilization response observed in Rhesus monkeys uponadministration of Gro-β T (450 μg/kg, intravenously) and plerixafor (1mg/kg, subcutaneously) Ratio of Ratio of fold Quantity of quantity ofincrease of cells observed CD34⁺ Fold increase CD34⁺ 4 hours Ratio ofCD90⁺ vs. baseline Ratio of fold CD90⁺ following iv quantity of CD45RA⁻following iv increase of CD45RA⁻ administration CD34⁺ cells cells toadministration CD34⁺ cells cells to other of Gro-beta T to other cellother cell of Gro-beta T to other cell cell Cell type at 450 μg/kg:populations populations at 450 μg/kg: populations populations CD34⁺47194 — 0.676 21.7 — 1.2 (38004- (0.393- (11.2-27.2) (1.1-4.8)  103119)0.745) CD34⁺ 25701 1.479 — 30.7 0.8 — CD90⁺ (16770- (1.341- (18.3-73.9)(0.2-0.9) CD54RA⁻ 76870) 2.545) WBCs 3.84E+07 0.0014 0.0009 3.8 5.1 6.0(2.72-5.27E+07) (0.0008- (0.0003- (2.7-5.4) (3.4-6.9) (5.5-26.9) 0.0021)0.0016) Neutrophils 1.79E+07 0.0036 0.0024 3.4 6.4 8.2 (1.03-2.44E+07)(0.0018- (0.0007- (3.1-5.2) (2.1-8.1) (3.5-22.0) 0.0058) 0.0043)Lymphocytes 1.76E+07 0.0031 0.0021 3.2 5.7 9.3 (0.8-2.39E+07) (0.0021-(0.0008- (2.0-5.4) (4.8-8.4) (5.6-37.0) 0.0094) 0.0069) Monocytes5.48E+06 0.0118 0.0073 11.7 1.6 1.9 (3.54-5.99E+06) (0.0071- (0.0028-(8.7-21.1) (1.1-2.3) (1.5-8.5)  0.0174) 0.0130)

TABLE 10 Mobilization response observed in Rhesus monkeys uponadministration of Gro-β T (450 μg/kg, subcutaneously and plerixafor (1mg/kg, subcutaneously) Ratio of Ratio of fold Quantity of quantity ofincrease of cells observed CD34⁺ Fold increase CD34⁺ 6 hours Ratio ofCD90⁺ vs. baseline Ratio of fold CD90⁺ following sc quantity of CD45RA⁻following sc increase of CD45RA⁻ administration CD34⁺ cells cells toadministration CD34⁺ cells cells to other of Gro-beta T to other cellother cell of Gro-beta T to other cell cell Cell type at 450 μg/kg:populations populations at 450 μg/kg: populations populations CD34⁺41178 — 0.359 6.3 — 1.0 (19413- (0.318- (4.8-13.2) (1.0-1.3) 72140)0.441) CD34⁺ 14782 2.786 — 6.6 1.0 — CD90⁺ (6177- (2.266- (6.1-13.6)(0.8-1.0) CD54RA⁻ 31841) 3.143) WBCs 6.31E+07 0.0007 0.0003 5.4 1.3 1.4(5.83-6.88E+07) (0.0003- (0.0001- (4.8-5.8)  (0.9-2.3) (1.1-2.3) 0.0011)0.0005) Neutrophils 4.00E+07 0.0011 0.0004 5.6 1.3 1.4 (3.64-4.98E+07)(0.0004- (0.0001- (4.8-6.1)  (0.8-2.4) (1.0-2.4) 0.0018) 0.0008)Lymphocytes 1.85E+07 0.0022 0.0008 4.5 1.4 1.6 (1.57-1.97E+07) (0.0012-(0.0004- (3.9-5.7)  (1.2-2.3) (1.5-2.4) 0.0037) 0.0016) Monocytes2.23E+06 0.0185 0.0066 9.7 0.7 0.8 (2.05-2.60E+06) (0.0075- (0.0024-(7.4-17.1) (0.6-0.8) (0.7-0.8) 0.0352) 0.0155)

TABLE 11 Mobilization response observed in Rhesus monkeys uponadministration of Gro-β T (1.2 mg/kg, subcutaneously) and plerixafor (1mg/kg, subcutaneously) Ratio of Ratio of fold Quantity of quantity ofincrease of cells observed CD34⁺ Fold increase CD34⁺ 4 hours Ratio ofCD90⁺ vs. baseline Ratio of fold CD90⁺ following sc quantity of CD45RA⁻following sc increase of CD45RA⁻ administration CD34⁺ cells cells toadministration CD34⁺ cells cells to other of Gro-beta T to other cellother cell of Gro-beta T to other cell cell Cell type at 1.2 mg/kg:populations populations at 1.2 mg/kg: populations populations CD34⁺36219 — 0.449 12.0 — 1.3 (26331- (0.278- (7.2-15.0) (0.8-1.9) 71704)0.548) CD34⁺ 17471 2.232 — 13.1 0.8 — CD90⁺ (7645- (1.826- (10.3-23.9) (0.5-1.3) CD54RA⁻ 25964) 3.594) WBCs 4.94E+07 0.0008 0.0003 5.7 1.8 2.5(4.07-6.66E+07) (0.0005- (0.0001- (4.5-9.7)  (1.4-2.7) (1.1-4.2) 0.0011)0.0005) Neutrophils 3.23E+07 0.0013 0.0006 9.7 1.2 1.6 (2.31-3.52E+07)(0.0008- (0.0002- (5.5-15.1) (0.7-1.9) (0.7-2.6) 0.0023) 0.0008)Lymphocytes 1.36E+07 0.0024 0.0010 3.3 3.6 4.9 (0.66-2.91E+07) (0.0019-(0.0005- (1.7-5.2)  (2.4-8.9)  (2.0-12.0) 0.0068) 0.0037) Monocytes4.08E+06 0.0130 0.0052 13.7 0.8 1.1 (1.16-5.37E+06) (0.0068- (0.0029-(7.2-23.2) (0.5-2.1) (0.6-2.8) 0.0237) 0.0078)

Conclusions

A single treatment of Gro-β T, in combination with AMD3100, inducesrobust mobilization of stem and progenitor cells within four hours ofadministration in nonhuman primates. Additionally, Gro-β T, incombination with AMD3100, results in 2-3 fold more CD34⁺ CD90⁺ CD45RA⁻stem and progenitor cells relative to AMD3100 alone, suggesting asignificant graft quality improvement.

Further, as evidenced by these data, Gro-β T, in combination withAMD3100, may offer a more robust and safer alternative to G-CSF inautologous and allogeneic transplant, including diseases such as sicklecell disease (SCD) and multiple sclerosis (MS) where G-CSF iscontraindicated or associated with adverse events.

Example 2. Determining Whether a Population of Hematopoietic Stem CellsMobilized with a CXCR2 Agonist and/or a CXCR4 Antagonist is Suitable forEx Vivo Expansion and/or Therapeutic Use

Using the compositions and methods described herein, a practitioner ofskill in the art may mobilize a population of hematopoietic stem orprogenitor cells in a mammalian donor, such as a human donor. In someembodiments, the practitioner may administer a CXCR4 antagonist and aCXCR2 agonist in amounts sufficient to engender the release of apopulation of a population of hematopoietic stem cells into circulatingperipheral blood while reducing the mobilization of other cells of thehematopoietic lineage, such as leukocytes, neutrophils, lymphocytes, andmonocytes.

When administering a CXCR4 antagonist in combination with a CXCR2agonist, the physician may administer the two agents to the donorsimultaneously or at different times. In some embodiments, the CXCR4antagonist may be administered to the donor from about 30 minutes toabout 180 minutes prior to administration of the CXCR2 agonist, such asfrom about 40 minutes to about 160 minutes, about 50 minutes to about150 minutes, about 60 minutes to about 140 minutes, about 70 minutes toabout 130 minutes, about 60 minutes to about 120 minutes, about 70minutes to about 110 minutes, or about 80 minutes to about 100 minutes(e.g., about 30 minutes, about 35 minutes, about 40 minutes, about 45minutes, about 50 minutes, about 55 minutes, about 60 minutes, about 65minutes, about 70 minutes, about 75 minutes, about 80 minutes, about 85minutes, about 90 minutes, about 95 minutes, about 100 minutes, about105 minutes, about 110 minutes, about 115 minutes, about 120 minutes,about 125 minutes, about 130 minutes, about 135 minutes, about 140minutes, about 145 minutes, about 150 minutes, about 155 minutes, about160 minutes, about 165 minutes, about 170 minutes, about 175 minutes, orabout 180 minutes prior to administration of the CXCR2 agonist).

To assess the efficacy of the mobilization regimen, a peripheral bloodsample may be isolated from the subject following administration of theCXCR2 agonist and/or CXCR4 antagonist. The sample may then becharacterized, for example, by acquiring an input value for each of oneor more parameters of the sample, such as a parameter listed in TABLE 2.Exemplary parameters that may be used to assess the efficacy of thehematopoietic stem cell mobilization regimen are ratios of hematopoieticstem cells to cells of other types, such as leukocytes, neutrophils,lymphocytes, and monocytes, as well as the relative frequency ofhematopoietic stem cells in the sample. Input values for theseparameters may be acquired, for example, using immunophenotyping methodsknown in the art, such as flow cytometry and fluorescence activated cellsorting (FACS) techniques.

When acquiring and analyzing input values for more than one parameterlisted in TABLE 2, one may analyze a combination of parameters. In someembodiments, one may analyze a ratio of hematopoietic stem cells toleukocytes, a ratio of hematopoietic stem cells to neutrophils, a ratioof hematopoietic stem cells to lymphocytes, a ratio of hematopoieticstem cells to monocytes, and/or the relative frequency of hematopoieticstem cells in a sample obtained from the peripheral blood of a donorfollowing administration of a CXCR2 agonist and/or a CXCR4 antagonist.One may analyze, for example, a combination of parameters set forth inany one of TABLES 3-6.

Upon acquiring an input value for each of the one or more parameters,one may then compare the input value(s) to the reference criterion foreach parameter. If the reference criterion is satisfied (e.g., if theratio of hematopoietic stem cells to another hematopoietic cell type issufficiently high, or if the relative frequency of hematopoietic stemcells in the sample obtained from the peripheral blood of the donor issufficiently high), then the cells may be released for ex vivo expansionand/or for therapeutic use.

Example 3. Treatment of a Hematologic Disorder by Administration of aHematopoietic Stem or Progenitor Cell Graft

Using the compositions and methods described herein, a practitioner ofskill in the art may treat a stem cell disorder, such as a hematologicpathology described herein, by administering to a patient ahematopoietic stem or progenitor cell graft. For example, a practitionermay identify a human donor of hematopoietic stem or progenitor cells aslikely to respond to a mobilization regimen, as outlined in Example 1,and may subsequently mobilize a population of hematopoietic stem orprogenitor cells accordingly, for example, as set forth in Example 2.Following mobilization, the physician may isolate a population ofhematopoietic stem or progenitor cells from the donor. Isolation of thecells may commence, for example, from about 10 minutes to about 60minutes following completion of the administration of a CXCR4 antagonistand/or a CXCR2 agonist, such as from about 15 minutes to about 55minutes, about 20 minutes to about 50 minutes, about 25 minutes to about45 minutes, or about 30 minutes to about 40 minutes following completionof the administration of these agents (e.g., about 10 minutes, about 15minutes, about 20 minutes, about 25 minutes, about 30 minutes, about 35minutes, about 40 minutes, about 45 minutes, about 50 minutes, about 55minutes, or about 60 minutes following completion of the administrationof the CXCR4 antagonist and the CXCR2 agonist).

The isolation procedure may be carried out over a period of from about15 minutes to about 6 hours, such as from about 20 minutes to about 4.5hours, about 30 minutes to about 4 hours, about 40 minutes to about 3.5hours, about 50 minutes to about 3 hours, or about 1 hour to about 2hours (e.g., over a period of about 15 minutes, about 20 minutes, about30 minutes, about 35 minutes, about 40 minutes, about 45 minutes, about50 minutes, about 55 minutes, about 60 minutes, about 65 minutes, about70 minutes, about 75 minutes, about 80 minutes, about 85 minutes, about90 minutes, about 95 minutes, about 100 minutes, about 105 minutes,about 110 minutes, about 115 minutes, or about 120 minutes). In someembodiments, the population of hematopoietic stem and progenitor cellsmay be isolated from the donor over a period of from about 30 minutes toabout 1 hour (e.g., over a period of about 30 minutes, about 35 minutes,about 40 minutes, about 45 minutes, about 50 minutes, about 55 minutes,or about 60 minutes).

Following the isolation process, a patient may then receive an infusion(e.g., an intravenous infusion) of the mobilized and isolatedhematopoietic stem or progenitor cells. The patient may be the donor, ormay be a patient that is HLA-matched with respect to the donor, therebyreducing the likelihood of graft rejection. The patient may be one thatis suffering, for example, from a cancer, such as a hematologic cancerdescribed herein. Additionally or alternatively, the patient may be onethat is suffering from an autoimmune disease or metabolic disorderdescribed herein. The physician may administer the patient the mobilizedand isolated hematopoietic stem or progenitor cells, for example, at adosage of from 1×10³ to 1×10⁹ hematopoietic stem cells/kg (e.g., about1×10⁵ CD34⁺ cells/kg to about 1×10⁷ CD34⁺ cells/kg, about 2×10⁵ CD34⁺cells/kg to about 9×10⁶ CD34⁺ cells/kg, about 3×10⁵ CD34⁺ cells/kg toabout 8×10⁶ CD34⁺ cells/kg, about 4×10⁵ CD34⁺ cells/kg to about 7×10⁶CD34⁺ cells/kg, depending on various factors, such as the patient's age,weight, and the severity of the disease being treated.

The physician may monitor the engraftment of the hematopoietic stem celltransplant, for example, by withdrawing a blood sample from the patientand determining the increase in concentration of hematopoietic stemcells or cells of the hematopoietic lineage (such as megakaryocytes,thrombocytes, platelets, erythrocytes, mast cells, myeoblasts,basophils, neutrophils, eosinophils, microglia, granulocytes, monocytes,osteoclasts, antigen-presenting cells, macrophages, dendritic cells,natural killer cells, T-lymphocytes, and B-lymphocytes) followingadministration of the transplant. This analysis may be conducted, forexample, from about 1 hour to about 6 months, from about 2 hours toabout 5 months, from about 3 hours to about 4 months, from about 4 hoursto about 3 months, from about 10 hours to about 7 days, from about 24hours to about 96 hours, or more following hematopoietic stem celltransplant therapy (e.g., 1 hour, 2 hours, 3 hours, 4 hours, 5 hours, 6hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 13hours, 14 hours, 15 hours, 16 hours, 17 hours, 18 hours, 19 hours, 20hours, 21 hours, 22 hours, 23 hours, 24 hours, 2 days, 3 days, 4 days, 5days, 6 days, 7 days, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 7weeks, 8 weeks, 9 weeks, 10 weeks, 11 weeks, 12 weeks, 13 weeks, 14weeks, 15 weeks, 16 weeks, 17 weeks, 18 weeks, 19 weeks, 20 weeks, 21weeks, 22 weeks, 23 weeks, 24 weeks, or more). A finding that theconcentration of hematopoietic stem cells or cells of the hematopoieticlineage has increased (e.g., by 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%,20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 500%, or more)following the transplant therapy relative to the concentration of thecorresponding cell type prior to transplant therapy provides oneindication that the hematopoietic stem or progenitor cell transplanttherapy is efficacious in treating the stem cell disorder.

Example 4: Mobilization of CD34^(dim) Cells Using a CXCR2 Agonist and/ora CXCR4 Antagonist

In this example, data are presented to demonstrate that combinationtreatment with Gro-β T and AMD3100 (plerixafor) results in significantlyenhanced mobilization of CD34^(dim) cells compared to that achieved withAMD3100 alone in nonhuman primates (NHP).

Cells were mobilized according to the method of Example 1. A sample ofperipheral blood was tested using a flow cytometer for the markers CD34,CD11b, CD14, CD16, CD45, and SSC.

As shown in FIG. 15, intravenous administration of 450 μg/kg Gro-β T andsubcutaneous administration of 1 mg/kg AMD3100 surprisingly leads to themobilization of a population of cells that are CD34^(dim) CD11b⁺.Subpopulations within this population are CD14⁺ CD16⁺, CD14⁻ CD16⁻, andcombinations thereof. FIG. 16 shows that mobilization of the CD34^(dim)cells is significantly enhanced upon administration of the combinationof Gro-β T and AMD3100 as compared to AMD3100 alone.

Example 5: Gro-β T in Combination with AMD3100 MobilizesImmunosuppressive Monocytes that Inhibit Graft Vs. Host Disease

This example demonstrates that combination treatment with Gro-β T andAMD3100 (plerixafor) results in significantly enhanced mobilization ofCD34′¹⁶⁶ cells having immunosuppressive properties and thattransplantation of this population of cells leads to a significantlyreduced incidence of acute graft versus host disease (aGVHD) in axenograft GVHD mouse model, as compared to transplantation of peripheralblood mononuclear cells having a lower concentration of CD34^(dim)cells.

Rhesus macaques were treated with Gro-β T, AMD310C, Gro-β T and AMD3100,or G-CSF alone. A sample of peripheral blood was tested using a flowcytometry. Animals mobilized with Gro-β T and AMD3100 showed a more thanfifteen-fold increase over baseline in the number of CD34^(dim) cells at4 hours post-treatment (p<0.0001, n=13, FIG. 17). By comparison, animalsmobilized with G-CSF alone showed only about a nine-fold increase overbaseline in the number of CD34^(dim) cells at 4 hours after the lastdose of G-CSF (p<0.0001, n=3, FIG. 17).

Further, the composition of unmobilized cells and grafts mobilized byG-CSF, Gro-β T and AMD3100 and AMD3100 alone is provided in FIG. 18. Asshown, grafts mobilized using Gro-β T and AMD3100 show a 3 fold increasein CD34^(dim) cells and a 3 fold increase in T-cells as compared tografts mobilized using G-CSF.

To determine if CD34^(dim) cells had immunosuppressive properties, theywere sorted from the peripheral blood of Rhesus macaques treated withGro-β T and AMD3100. The CD34^(dim) cells were co-cultured withcarboxyfluorescein succinimidyl ester (CFSE)-labeled autologous T cellsstimulated with anti-CD2/CD3/CD28-coated beads in vitro. As shown inFIG. 19, Gro-β T and AMD3100 mobilized CD34^(dim) cells suppressedT-cell proliferation as measured by CFSE staining after four days.

To assess whether these immunosuppressive cells may prevent GVHD, wedeveloped a xenograft GVHD model in NOD scid gamma (NSG) mice, which areimmunodeficient mice. Gro-β T and AMD3100 mobilized peripheral blood(6×10⁶ PBMCs) containing a high percentage of CD34^(dim) cells wereinjected into sublethally irradiated NSG mice. This was compared tounmobilized primate PBMCs (6×10⁶ PBMCs) containing a relatively lownumbers of CD34^(dim) cells and G-CSF mobilized primate PBMCs (6×10⁶PBMCs). As shown in the survival curve in FIG. 20, at day 24, all mice(13/13) transplanted with unmobilized PBMCs had died of aGVHD comparedto 5/16 mice transplanted with AMD3100 mobilized PBMCs, 3/16 micetransplanted with G-CSF-mobilized PBMCs and none of the micetransplanted with Gro-β T and AMD3100-mobilized PBMCs. At day 60post-transplant, 15/16 mice transplanted with Gro-β T and AMD3100remained alive, whereas only 10/16 mice transplanted with AMD3100mobilized peripheral blood and 11/16 mice transplanted with G-CSFmobilized peripheral blood remained alive (see FIG. 20). As shown inFIG. 21A, mice transplanted with unmobilized PBMCs (“Unmobilized”) alsodemonstrated higher numbers of donor T-cells (approximately 160-foldhigher, see FIG. 21A) compared to mice transplanted with Gro-β T andAMD3100 mobilized PBMCs (“Gro-β T+Plerixafor”) at day 14 post-transplant(*p<0.01, n=6-8). Additionally, also as shown in FIG. 21A, micetransplanted with PBMCs mobilized with AMD3100 (i.e., plerixafor) alone(“Plerixafor”) or PBMCs mobilized with G-CSF alone (“G-CSF”)demonstrated higher numbers of T-cells (approximately 60-fold higher(p<0.05) and approximately 10-fold higher (p<0.01), respectively) ascompared to mice transplanted with Gro-β T and AMD3100-mobilized PBMCs.

To confirm that the CD34^(dim) cells in the Gro-β T andAMD3100-mobilized graft were responsible for the protection againstaGvHD in the NSG mouse xenotransplantation model shown in FIG. 20,additional cohorts of mice were transplanted with 6×10⁶ PBMCs from Gro-βT and AMD3100-mobilized rhesus monkeys, where CD34^(dim) cells werefirst depleted by fluorescence activated cell sorting and T cell numbersand survival were compared to cohorts transplanted with Gro-β T andAMD3100-mobilized PBMCs and cohorts transplanted with unmobilized PBMCs.As shown in FIG. 21B, at day 14 post-transplant, mice transplanted withGro-β T and AMD3100-mobilized PBMCs (“Gro-β T+Plerixafor”) hadsignificantly reduced numbers of rhesus CD45+CD3+ T cells in peripheralblood as compared to mice transplanted with unmobilized PBMCs(“Umnobilized”) (p<0.001, n=10/group). By contrast, mice transplantedwith Gro-β T and AMD3100 mobilized PBMCs that were depleted of CD34dimcells showed no significant difference in T-cell numbers by day 14post-transplant (n=10) as compared to mice transplanted with unmobilizedPBMCs (“Unmobilized”) (see FIG. 21B). As shown in FIG. 21C, by day 44post transplant, 0/10 mice transplanted with unmobilized PBMCs(“Unmobilized”) survived versus 8/10 mice transplanted with Gro-β T andAMD3100-mobilized PBMCs (“Gro-β T+plerixafor”) (p<0.05) and only 4/10mice transplanted with Gro-β T and AMD3100 mobilized PBMCs that weredepleted of CD34dim cells (“Gro-β T+plerixafor CD34^(dim)-depleted”) (nosignificant difference versus Unmobilized).

Accordingly, these data demonstrate that co-administration of Gro-β Tand AMD3100 results not only in rapid and efficacious mobilization ofhighly enriched HSCs but that these HSCs are enriched in a CD34^(dim)cell population with potent immunosuppressive activity compared toAMD3100 alone or the current standard of care, G-CSF, and thattransplant of this CD34^(dim) cell population can result in a reducedincidence of or risk for aGVHD.

Other Embodiments

All publications, patents, and patent applications mentioned in thisspecification are incorporated herein by reference to the same extent asif each independent publication or patent application was specificallyand individually indicated to be incorporated by reference.

While the invention has been described in connection with specificembodiments thereof, it will be understood that it is capable of furthermodifications and this application is intended to cover any variations,uses, or adaptations of the invention following, in general, theprinciples of the invention and including such departures from theinvention that come within known or customary practice within the art towhich the invention pertains and may be applied to the essentialfeatures hereinbefore set forth, and follows in the scope of the claims.

Other embodiments are within the claims.

1-167. (canceled)
 168. A method of performing an allogeneichematopoietic stem cell transplant in a patient in need thereof, themethod comprising infusing into the patient a therapeutically effectiveamount of allogeneic hematopoietic stem cells, wherein the hematopoieticstem cells were mobilized from bone marrow of a human donor intoperipheral blood of the human donor by a method comprising administeringto the donor (i) a CXCR2 agonist selected from the group consisting ofGro-β, Gro-β T, and variants thereof at a dose of from about 50 μg/kg toabout 1,000 μg/kg and (ii) a CXCR4 antagonist, wherein CD34^(dim) cellsare present in the peripheral blood in an amount that is at least two-to ten-fold higher than if the hematopoietic stem cells were mobilizedusing the CXCR4 antagonist alone.
 169. The method of claim 168, whereinthe CD34^(dim) cells are capable of suppressing alloreactive Tlymphocyte proliferation when administered to a recipient.
 170. Themethod of claim 168, wherein the CXCR2 agonist is Gro-β T.
 171. Themethod of claim 168, wherein the CXCR2 agonist is administered to thedonor at a dose of from about 100 μg/kg to about 250 μg/kg.
 172. Themethod of claim 171, wherein the CXCR2 agonist is administered to thedonor at a dose of from about 125 μg/kg to about 225 μg/kg.
 173. Themethod of claim 172, wherein the CXCR2 agonist is administered to thedonor at a dose of about 150 μg/kg.
 173. The method of claim 168,wherein the CXCR2 agonist is administered intravenously to the donor.174. The method of claim 168, wherein the CXCR4 antagonist isadministered subcutaneously to the donor.
 175. The method of claim 168,wherein the CXCR4 antagonist is plerixafor or a pharmaceuticallyacceptable salt thereof.
 176. The method of claim 175, wherein theplerixafor or pharmaceutically acceptable salt thereof is administeredto the donor at a dose of from about 50 μg/kg to about 500 μg/kg. 177.The method of claim 176, wherein the plerixafor or pharmaceuticallyacceptable salt thereof is administered to the donor at a dose of fromabout 200 μg/kg to about 300 μg/kg.
 178. The method of claim 177,wherein the plerixafor or pharmaceutically acceptable salt thereof isadministered to the donor at a dose of about 240 μg/kg.
 179. The methodof claim 168, wherein the method prevents, reduces the risk ofdeveloping, or reduces the severity of a post-transplant infection inthe patient.
 180. The method of claim 168, wherein the method prevents,reduces the risk of developing, or reduces the severity of graft versushost disease (GVHD) in the patient.